CN1173989C - DNA sequence of dendrobium and method for discriminating its veriety and judging if it is true or false - Google Patents

DNA sequence of dendrobium and method for discriminating its veriety and judging if it is true or false Download PDF

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CN1173989C
CN1173989C CNB011024348A CN01102434A CN1173989C CN 1173989 C CN1173989 C CN 1173989C CN B011024348 A CNB011024348 A CN B011024348A CN 01102434 A CN01102434 A CN 01102434A CN 1173989 C CN1173989 C CN 1173989C
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its2
cgc
dendrobium
gtc
ttt
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CN1368504A (en
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邵鹏柱
王骏
毕培曦
刘大伟
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Chinese University of Hong Kong CUHK
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Chinese University of Hong Kong CUHK
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Abstract

The present invention discloses partial DNA sequences of ITS2 ribosome, 5.8S ribosome and 28S ribosome for 16 kinds of dendrobe and one kind of Chinese pholidota herb. The DNA sequences are obtained by PCR amplification reaction and a direct sequence measuring method and can be used to discriminate the variety of dendrobe and to judge the reality of the dendrobe.

Description

The dna sequence dna of the stem of noble dendrobium and utilize its kind of this Sequence Identification or distinguish the method for its true and false
The present invention relates to the dna sequence dna of valuable Chinese medicine and utilize its kind of this Sequence Identification and the method for distinguishing its true and false, more particularly, the present invention relates to the ITS2 of the Chinese medicine stem of noble dendrobium and 5.8S and 28S rrna dna sequence dna and utilize its kind of this Sequence Identification or distinguish the method for its true and false partly partly.
The stem of noble dendrobium is valuable Chinese medicinal materials, and applicating history is long, classifies as top gradely in Shennong's Herbal, has enriching yin disappear heat, the beneficial stomach that promotes the production of body fluid, effect such as moisten the lung and relieve the cough, and is used for consumption of body fluid caused by febrile disease, dry polydipsia, various disease conditions such as Virtual heat after being ill.Modern pharmacological research shows, the effect that the stem of noble dendrobium also has is antitumor, anti-ageing, strengthen body immunity and vasodilation.Therefore the stem of noble dendrobium reaches in the Chinese traditional compound medicine clinically and is widely used.Chinese Pharmacopoeia (nineteen ninety-five version) has recorded 5 kinds of stems of noble dendrobium, be dendrobium stem (D.nobile Lindl.), Herba Dendrobii (D.candidum Wall.ex Lindl.), Dendrobium chrysanthum (D.chrysanthum Wall.ex.Lindl.), the U.S. flower stem of noble dendrobium (D.loddigesii Rolfe) and Dendrobium fimbriatum Hook. (D.fimbriatum Hook).
The market demand of stem of noble dendrobium medicinal material is very big, and reach more than the thousands of tons of every year, and the fetch long price of part commodity, for example the price of the one-level stem of noble dendrobium (Herba Dendrobii) per kilogram reaches 3500 dollars.
The source complexity of the commercially available stem of noble dendrobium relates to Dendrobium plant kind more than 30, and the time have the orchid (belonging to) of other genus to mix up stem of noble dendrobium medicinal material to use, directly have influence on the clinical efficacy of the stem of noble dendrobium and retail consumer's interests as Pseudobulbus Pholidotae Chinensis.Moreover the former plant of stem of noble dendrobium medicinal material all is endangered breeds, lists in the III class level Do of Chinese protected plant register as Herba Dendrobii.
Existing stem of noble dendrobium authentication method comprises proterties Jian Do, chemical analysis and microcosmic dissection.But these methods are not done to identify to the listed stem of noble dendrobium kind of pharmacopeia yet comprehensively and objectively.
Plant ribosome thymus nucleic acid (rDNA) exists with the form of the anti-sub-18S-ITS1-5.8S-ITS2-26S of Cis.Plant has the tumor-necrosis factor glycoproteins of the anti-son of this Cis mostly, and it accumulates on the karyomit(e) in mode from beginning to end.Studies show that at present the base-pair sequence of rDNA genes such as 18S, 5.8S and 26S is very conservative, De Cha Do is very little between different plants; The then poor Do of base-pair sequence of ITS1 between plant rDNA gene (transcribing interstice coverage 1) and ITS2 (transcribing interstice coverage 2) is bigger.Simultaneously because the effect of gene transformation and exchange, with regard to same product kind of plant, its rDNA sequence roughly the same (Hillis and Dixon, 1991).
Therefore, the ITS2 and 5.8S and the 28S rrna part dna sequence dna that the purpose of this invention is to provide stem of noble dendrobium medicinal material.
Another object of the present invention provides the kind of utilizing the described dna sequence dna evaluation stem of noble dendrobium and the method for distinguishing its true and false.
To achieve the object of the present invention, the inventor from Hong Kong and the different location of China's Mainland collected 46 kinds of samples, comprise 16 kinds of stems of noble dendrobium and a kind of Pseudobulbus Pholidotae Chinensis.All samples all pass through the orchid family expert statement.Except the stem of noble dendrobium of sieve river, all the other each kinds all have the sample that obtains from several different groups.Partly dna sequence dna is as follows for the ITS2 of 46 measured samples and 5.8S and 28S rrna:
1. the ITS2 of hooked dendrobium (Dendrobium aduncum Lindl.) sample 1 and 5.8S and 28S rrna part rDNA sequence:
AACCATCG?AGTCTTTG?AACGCAAG?TTGCGCCC?GAGGCCAA
CCGGCTAA?GGGCACGT?CCGCCTGG?GCGTCAAG?CATTTTAT
CGCTCCGT?GCCTAATC?TCCCATCC?ATGTATGT?GTTGCAAA
GGCTCGGA?TGTGCACG?GTGGCTCC?TCGTGCCC?CTTGGTGC
GGCGGGCT?GAAGGCCG?GGTCATCT?TCTCGTTG?GTTGCCAA
CAATAAGG?GGTGGATT?AAACAAGG?CCTATGCT?GTTGTGAT
ATGCGCGC?CCGAGAGA?TGATCATA?CTTTTTAG?GTGATCCC
AATTCATG?CGCTGATC?CATGGATG?GCGTATCG?AATGCGAC
CCCAGGAT?GGGCGAGG?CCACCCGC?TGAGTTTA?AGCATATC
AATAAGCG?GAGGAAAA?GAAACTTA?CGAGGATT?CCCCTAGT
AACGGCGA?GT
2. the ITS2 of hooked dendrobium (Dendrobium aduncum Lindl.) sample 2 and 5.8S and 28S rrna rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCGAGGCC
AACCGGCT?AAGGGCAC?GTCCGCCT?GGGCGTCA?AGCATTTT
ATCGCTCC?GTGCCTAA?TCTCCCAT?CCATGTAT?GTGTTGCA
AAGGCTCG?GATGTGCA?CGGTGGCT?CCTCGTGC?CCCTTGGT
GCGGCGGG?CTGAAGGC?CGGGTCAT?CTTCTCGT?TGGCTGCC
AACAATAA?GGGGTGGA?TTAAACAA?GGCCTATG?CTGTTGTG
ATATGCGC?GCCCGAGA?GATGATCA?TACTTTTT?AGGTGATC
CCAATTCA?TGCGCTGA?TCCATGGA?TGGCGTAT?CGAATGCG
ACCCCAGG?ATGGGCGA?GGCCACCC?GCTGAGTT?TAAGCATA
TCAATAAG CGGAGG hook AAAAGAAA CTTACGAG GATTCCCC
TAGTAACG?GCGAGT
3. the ITS2 of hooked dendrobium (Dendrobium aduncum Lindl.) sample 3 and 5.8S and 28S rrna rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCGAGGCC
AACCGGCT?AAGGGCAC?GTCCGCCT?GGGCGTCA?AGCATTTT
ATCGCTCC?GTGCCTAA?TCTCCCAT?CCATGTAT?GTGTTGCA
AAGGCTCG?GATGTGCA?CGGTGGCT?CCTCGTGC?CCCTTGGT
GCGGCGGG?CTGAAGGC?CGGGTCAT?CTTCTCGT?TGGTTGCC
AACAATAA?GGGGTGGA?TTAAACAA?GGCCTATG?CTGTTGTG
ATATGCGC?GCCCGAGA?GATGATCA?TACTTTTT?AGGTGATC
CCAATTCA?TGCGCTGA?TCCATGGA?TGGCGTAT?CGAATGCG
ACCCCAGG?ATGGGCGA?GGCCACCC?GCTGAGTT?TAAGCATA
TCAATAAG?CGGAGGAA?AAGAAACT?TACGAGGA?TTCCCCTA
GTAACGGC?GAGT
4. the ITS2 of Dendrobium chrysanthum (Dendrobium chrysanthum Lindl.) sample 1 and 5.8S and 28S rrna rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCCGAGGC
CAACCTGC?CAAGGGCA?CGTCTGCC?TGGGCGTC?AAGCATTT
TGTTGCTC?CGTGTCTC?GTCACCCA?TCAATGGA?TGTGCTGG
CGAAGGCT?CGGATGTG?CACGGTGG?CCCGTCGT?GCCCCTCG
GTGCGGCG?GGCTTAAG?AGCGGGTC?GCCATCTC?GTTGGTTG
CGAACAAT?AAGGGGTG?GATTAAAT?GGAGGCCT?ATGCTATT
GTGTCGTG?CATGCCTA?AGAGATTAAA?TTTTTTAG?GTGATCCC
AAATCACG?CGTCGATC?CACGGATG?ATGCTTTG?AATGCGAC
CCCAGGAT?GGGCGAGG?CCACCCGC?CGAGTTTA?AGCATATC
AATAAGCG?GAGGAGAA?GAAACTTA?CAAGGATT?CCCCTAGT
AACGGCGA?GT
5. the ITS2 of Dendrobium chrysanthum (Dendrobium chrysanthum Lindl.) sample 2 and 5.8S and 28S rrna rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AA?GTT?GCG?CCCGAGGC
CAACCTGC?CAAGGGCA?CGTCTGCC?TGGGCGTC?AAGCATTT
TGTTGCTC?CGTGTCTC?GTCACCCA?TCAATGGA?TGTGCTGG
CGAAGGCT?CGGATGTG?CACGGTGG?CCCGTCGT?GCCCCTCG
GTGCGGCG?GGCTTAAG?AGCGGGTC?GCCATCTC?GTTGGTTG
CGAACAAT?AAGGGGTG?GATTAAAT?GGAGGCCT?ATGCTATT
GTGTCGTG?CATGCCTA?AGAGATTA?AATTTTTT?AGGTGATC
CCAAATCA?CGCGTCGA?TCCACGGA?TGATGCTT?TGAATGCG
ACCCCAGG?ATGGGCGA?GGCCACCC?GCCGAGTT?TAAGCATA
TCAATAAG?CGGAGGAG?AAGAAACT?TACAAGGA?TTCCCCT
AGTAACGG?CGAGT
6. the ITS2 of Dendrobium chrysotoxum (Dendrobium chrysotoxum Lindl.) sample 1 and 5.8S and 28S rrna rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCGAGGCC
AATCGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGCGTTAC
GTCGCTCC?GTGTCCAG?CCACCCGT?CGATGGAT?GGGCTGGC
GATGGCTC?GGATGTGC?ACAGTGGC?TCCTCGTG?CCCCTCGG
CACGGCGG?GCTTAAGA?GCGGGTAA?TCATCTCG?TTGGCTGC
GAACAATA?TGGGGTGG?ATAAATGC?GATGCTTA?TGATATTG
TGCCGTGC?ATGCCCGA?GAGATGAT?CATACTTA?TTCAGGTG
ATCCCAAA?CCATGCGT?CGATCCAT?GGATGGCG?TTTTGAAT
GCGACCCC?AGGACAGG?CGAGGCCA?CCCGCCGA?GTTTAAGC
ATATCAAT?AAGCGGAG?GAGAAGAA?ACTTACGA?GGATTCCC
CTAGTAAC?GGCGAGT
7. the ITS2 of Dendrobium chrysotoxum (Dendrobium chrysotoxum Lindl.) sample 2 and 5.8S and 28S rrna rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCGAGGCC
AATCGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGCGTTAC
GTCGCTCC?GTGTCCAG?CCACCCGT?CGATGGAT?GGGCTGGC
GATGGCTC?GGATGTGC?ACAGTGGC?TCCTCGTG?CCCCTCGG
CACGGCGG?GCTTAAGA?GCGGGTAA?TCATCTCG?TTGGCTGC
GAACAATA?TGGGGTGG?ATAAATGC?GATGCCTA?TGATATTG
TGCCGTGC?ATGCCCGA?GAGATGAT?CATACTTA?TTCAGGTG
ATCCCAAA?CCATGCGT?CGATCCAT?GGATGGCG?TTTTGAAT
GCGACCCC?AGGACAGG?CGAGGCCA?CCCGCCGA?GTTTAAGC
ATATCAAT?AAGCGGAG?GAGAAGAA?ACTTACGA?GGATTCCC
CTAGTAAC?GGCGAGT
8. the ITS2 of Dendrobium chrysotoxum (Dendrobium chrysotoxum Lindl.) sample 3 and 5.8S and 28S rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCGAGGCC
AATCGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGGTTACG
TCGCTCCG?TGTCCAGC?CACCCGTC?GATGGATG?GGCTGGCG
ATGGCTCG?GATGTGCAC?AGTGGCTC?CTCGTGCC?CCTCGGCA
CGGCGGGC?TTAAGAGC?GGGTAATC?ATCTCGTT?GGCTGCGA
ACAATATG?GGGTGGAT?AAATGCGA?TGCTTATG?ATATTGTG
CCGTGCAT?GCCCGAGA?GATGATCA?TACTTATT?CAGGTGAT
CCCAAACC?ATGCGTCG?ATCCATGG?ATGGCGTT?TTGAATGC
GACCCCAG?GACAGGCG?AGGCCACC?CGCCGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?GAAGAAAC?TTACGAGG?ATTCCCCT
AGTAACGG?CGAGT
9. the ITS2 of Dendrobium crepidatum lindl et paxt (Dendrobium crepidatum Lindl.ex Paxt.) sample 1 and 5.8S and 28S rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCG?AGG
CCAATCGG?CCAAGGGC?ACGTCTGC?CTGGGCGT?CAAGCATT
TTGTTGCT?CCGTGTCT?ACTCACAC?CCATCGAT?GGATGTGC
TGGTGTAG?GCTCGGAT?GTGCATGC?TGGCTCGT?CGTTCTCC
CAGGTGCG?GCGGGCTG?AAGAGCGG?GTCATCCT?CTCGTTTG
CGGCGAAC?AATAAGGG?GTGGATTG?AAGAGAGG?TCTATGCT
ATTGTGTC?GTGCATGC?CTGAGAGA?TGATTAAC?TTTTTTGT
GATCCTAA?ATCATGCA?TCGATCCA?TGGATGAT?GCTTTGAA
TGTGACCC?CAGGATGG?GCGAGGCT?ACCCGCTG?AGTTTAAG
CATATCAA?TAAGCGGA?GGAGAAGA?AACTTACG?AGGATTCC
CCTAGTAA?CGGCGAGT
10. the ITS2 of Dendrobium crepidatum lindl et paxt (Dendrobium crepidatum Lindl.ex Paxt.) sample 2 and 5.8S and 28S rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCGAGGCC
AATCGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGCATTTT
GTTGCTCC?GTGTCTAC?TCACACCC?ATCGATGG?ATGTGCTG
GTGTAGGC?TCGGATGT?GCATGCTG?GCTCGTCG?TTCTCCCA
GGTGCGGC?GGGCTGAA?GAGCGGGT?CATCCTCT?CGTTTGCG
GCGAACAA?TAAGGGGT?GGATTGAA?GAGAGGTC?TATGCTAT
TGTGTCGT?GCATGCCT?GAGAGATG?ATTAACTT?TTTTGTGA
TCCCAAAT?CATGCATC?GATCCATG?GATGATGC?TTTGAATG
TGACCCCA?GGATGGGC?GAGGCTAC?CCGCTGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AGAAGAAA?CTTACGAG?GATTCCCC
TAGTAACG?GCGAGT
11. the ITS2 of Dendrobium crepidatum lindl et paxt (Dendrobium crepidatum Lindl.ex Paxt.) sample 3 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGCCCGAGGC
CAATCGGC?CAAGGGCA?CGTCTGCC?TGGGCGTC?AAGCATTT
TGTTGCTC?CGTGTCTA?CTCACACC?CATCGATG?GATGTGCT
GGTGTAGG?CTCGGATG?TGCATGCT?GGCTCGTC?GTTCTCCC
AGGTGCGG?CGGGCTGA?AGAGCGGG?TCATCCTC?TCGTTTGC
GGCGAACA?ATAAGGGG?TGGATTGA?AGAGAGGT?CTATGCTA
TTGTGTCG?TGCATGCC?TGAGAGAT?GATTAACT?TTTTTGTG
ATCCCAAA?TCATGCAT?CGATCCAT?GGATGATG?CTTTGAAT
GTGACCCC?AGGATGGG?CGAGGCTA?CCCGCTGA?GTTTAAGC
ATATCAAT?AAGCGGAG?GAGAAGAA?ACTTACGA?GGATTCC
CCTAGTAC?GGCGAGT
12. the ITS2 of dendrobium (Dendrobium densiflorum Lindl.) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?A?AG?TTG?CGC?CCG?AGG
CCAACCGG?CCAAGGGC?ACGTCTGC?CTGGGCGT?CAAGCGTT
ATGTCGCT?CCGTGTTA?ACTCCATC?CCATCGAT?GTATGGGC
TGGCGAAG?GCTCGGAT?GTGCAGTG?TGGCTCGT?CGTGCCCC
TTGGTGCG?GCGAGCTG?AAAAGCGG?GTAATCAT?CTCGTTGG
CTGCGAAC?GATAAGGG?GTGGATTA?AAGCGAGA?CCTATGTT
ATTGTGTC?GTGTATGC?CCGAGAGA?AGATTATA?CATACTCA
GGTGATCC?TGAATCAT?GCGTCGAT?CAAAGGAC?GGCGCTTG
GAATGCGA?CCCCAGGA?TGGGCGAG?GCCACCCG?CCGAGTTT
AAGCATAT?TAATAAGC?GGAGGAGA?AGAAACTT?ACGAGGAT
TCCCCTAG?TAACGGCG?AGT
13. the ITS2 of dendrobium (Dendrobium densiflorum Lindl.) sample 2 and 5.8S and 28S be the rDNA sequence partly:
AACCATCG?AGTCTTTGA?ACGCAAGT?TGCGCCCG?AGGCCAAC
CGGCCAAG?GGCACGTC?TGCCTGGG?CGTCAAGC?GTTATGTC
GCTCCGTG?TTAACTCC?ATCCCATC?GATGTATG?GGCTGGCG
AAGGCTCG?GATGTGCA?GTGTGGCT?CGTCGTGC?CCCTTGGT
GCGGCGAG?CTGAAAAG?CGGGTAAT?CATCTCGT?TGGCTGCG
AACGATAA?GGGGTGGA?TTAAAGCG?AGACCTAT?GTTATTGT
GTCGTGTA?TGCCCGAG?AGAAGATT?ATACATAC?TCAGGTGA
TCCTGAAT?CATGCGTC?GATCAAAG?GACGGCGC?TTGGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CCGCCGAG?TTTAAGCA
TATTAATA?AGCGGAGG?AGAAGAAA?CTTACGAG?GATTCCC?C
TAGTAACG?GCGAGT
14. the ITS2 of dendrobium (Dendrobium densiflorum Lindl.) sample 3 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCG?AGGCCA
ACCGGCCA?AGGGCACG?TCTGCCTG?GGCGTCAA?GCGTTATG
TCGCTCCG?TGTTAACT?CCATCCCA?TCGATGTA?TGGGCTGG
CGAAGGCT?CGGATGTG?CAGTGTGG?CTCGTCGT?GCCCCTTG
GTGCGGCG?AGCTGAAA?AGCGGGTA?ATCATCTC?GTTGGCTG
CGAACGAT?AAGGGGTG?GATTAAAG?CGAGACCT?ATGTTATT
GTGTCGTG?TATGCCCG?AGAGAAGA?TTATACAT?ACTCAGGT
GATCCTGA?ATCATGCG?TCGATCAA?AGGACGGC?GCTTGGAA
TGCGACCC?CAGGATGG?GCGAGGCC?ACCCGCCG?AGTTTAAG
CATATTAA?TAAGCGGA?GGAGAAGA?AACTTACG?AGGATTCC
CCTAGTAA?CGGCGAGT
15. the ITS2 of Dendrobium fimbriatum Hook. (Dendrobium fimbriatum Hook.) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AACCATCG?AGTCTTTG?AACGCAAG?TTGCGCCC?GAGGCCAA
CAGGCCAA?GGGCACGT?CTGCCTGG?GCGTCAAG?TGTTATGT
CGCTCCAT?GCCAAGTC?ACCCATCG?ATGGATGG?GCCGGCGA
AGGCTCGG?ATGTGCAC?AGTGGCTC?GCCGTGCC?CCTCGGTG
CGGCGGGC?TGAAGAGC?GGGTCATT?ATCTTGTT?GGCTGCAG
GCAATAAT?GGGTGGAT?AAACGCGA?GGCCTTTG?TTATTGTG
TCGTGCAT?GCCTAAGA?TATGATTA?AACTTTTT?AGGTGAT
CCCAAATC?ATGCGTCG?ATCCACGG?ATGGCGTT?TTGAATGC
GACCCCAG?GATGGGCG?AGGCCACC?CGCCGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?GAAGAAAC?TTACGAGG?ATTCCCCT
AGTAACGG?CGAGT
16. Dendrobium fimbriatum Hook. (Dendrobium fimbriatum Hook.) sample 2ITS2 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCGAGGCC
AACAGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGTGTTAT
GTCGCTCC?ATGCCAAG?TCACCCAT?CGATGGAT?GGGCCGGC
GAAGGCTC?GGATGTGC?ACAGTGGC?TCGCCGTG?CCCCTCGG
TGCGGCGG?GCTGAAGA?GCGGGTCA?TTATCTTG?TTGGCTGC
AGGCAATA?ATGGGTGG?ATAAACGC?GAGGCCT?TTGTTATT
GTGTCGTG?CATGCCTA?AGATATGA?TTAAACTT?TTTAGGTG
ATCCCAAA?TCATGCGT?CGATCCAC?GGATGGCG?TTTTGAAT
GCGACCCC?AGGATGGG?CGAGGCCA?CCCGCCGA?GTTTAAGC
ATATCAAT?AAGCGGAG?GAGAAGAA?ACTTACGA?GGATTCCC
CTAGTAAC?GGCGAGT
17. Dendrobium fimbriatum Hook. (Dendrobium fimbriatum Hook.) sample 3ITS2 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCGAGGCC
AACAGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGTGTTAT
GTCGCTCC?ATGCCAAG?TCACCCAT?CGATGGAT?GGGCCGGC
GAAGGCTC?GGATGTGC?ACAGTGGC?TCGCCGTG?CCCCTCGG
TGCGGCGG?GCTGAAGA?GCGGGTCA?TTATCTTG?TTGGCTGC
AGGCAATA?ATGGGTGG?ATAAACGC?GAGGCCTT?TGTTATTG
TGTCGTGC?ATGCCTAA?GATATGAT?TAAACTTT?TTAGGTGA
TCCCAAAT?CATGCGTC?GATCCACG?GATGGCGT?TTTGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CCGCCGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AGAAGAAA?CTTACGAG?GATTCCCC
TAGTAACG?GCGAGT
18. the ITS2 of the heavy lip stem of noble dendrobium (Dendrobium hercoglossum Rchb.f.) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AA?CCAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CG?CCTGAGGC
CAACCGGC?TGAGGGCA?CGTCCGCC?TGGGCGTC?AAGCATTT
TATCGCTC?CGTGCCTA?GTCTCCCA?TCCATGGA?TGTGTTGC
CAAGGCTC?GGATGTGC?ACGGTGGC?TCGTCGTG?CCCCTTGG
TGCGGCGG?GCTGAAGG?GCGGGTCA?TCTTCTCG?TTGGCTGC
CAACAATA?AGGGGTGG?ATTAAATA?AGGCCTAT?GCTATTGT
GTCAAGCG?CGCCTGAG?AGATGGTC?ATACTTAT?TAGGTGAT
CCCAATTC?ATGCGTCG?ATCCATGG?ATGGCGTA?TCGAATGT
GACCCCAG?GATGGGCG?AGGCCACC?CGCCGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?GAAGAAAC?TTACGAGG?ATTCCCCT
AGTAACGG?CGAGT
19. the ITS2 of the heavy lip stem of noble dendrobium (Dendrobium hercoglossum Rchb.f.) sample 2 and 5.8S and 28S be the rDNA sequence partly:
AACCAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CG?CCT?GAGGC
CAACCGGC?TGAGGGCA?CGTCCGCC?TGGGCGTC?AAGCATTT
TATCGCTC?CGTGCCTA?GTCTCCCA?TCCATGGA?TGTGTTGC
CAAGGCTC?GGATGTGC?ACGGTGGC?TCGTCGTG?CCCCTTGG
TGCGGCGG?GCTGAAGG?GCGGGTCA?TCTTCTCG?TTGGCTGC
CAACAATA?AGGGGTGG?ATTAAATA?AGGCCTAT?GCTATTGT
GTCAAGCG?CGCCTGAG?AGATGGTCA?TACTTATT?AGGTGATC
CCAATTCA?TGCGTCGA?TCCATGGA?TGGCGTAT?CGAATGTG
ACCCCAGG?ATGGGCGA?GGCCACCC?GCCGAGTT?TAAGCATA
TCAATAAG?CGGAGGAG?AAGAAACT?TACGAGGA?TTCCCCTA
GTAACGGC?GAGT
20. the ITS2 of the Xiao Yellow flower stem of noble dendrobium (Dendrobium jenkinsii Lindl.) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AACCATCG?AGTCTTTG?AACGCAAG?TTGCGCCC?GAGGCCAA
TCGGCCAA?GGGCACGT?TCGCCTGG?GCGTCAAG?CATTACGT
CGCTCCAT?GTCAACTC?CATCC?CTTCTTTTGA?TGGGCTGGT
AAAAGGCT?TGGACGTG?TAAAGTGG?CTCGTCGT?GCCCCTCG
GTGCGGCG?GGCTTAAG?AGCGGGTC?ATCATCTC?GTTGGCTA
CGAACAAT?TACGTGTG?GATTAAAG?TGCGGCCT?ATCTTATT
GTGTCATG?TATGCCTG?AGAAGATT?ATGCATTT?TTAGGTGA
TCCCAAAT?CATGCGTC?GATCCATG?GATGACGT?TTGGAATG
CGACCCCA?GGATGGGC?GAGGTCAC?CCGCTGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AGAAGAAA?CTTACGAG?GATTCCCC
TAGTAACG?GCGAGT
21. the ITS2 of the Xiao Yellow flower stem of noble dendrobium (Dendrobium jenkinsii Lindl.) sample 2 and 5.8S and 28S be the rDNA sequence partly:
AACCAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCGAGGCC
AATCGGCC?AAGGGCAC?GTTCGCCT?GGGCGTCA?AGCATTAC
GTCGCTCC?ATGTCAAC?TCCATCCC?TTCTTTTG?ATGGGCTG
GTAAAAGG?CTTGGACG?TGTAAAGT?GGCTCGTC?GTGCCCCT
CGGTGCGG?CGGGCTTA?AGAGCGGG?TCATCATC?TCGTTGGC
TACGAACA?ATTACGTG?TGGATTAA?AGTGCGGC?CTATCTTA
TTGTGTCA?TGTATGCC?TGAGAAGA?TTATGCAT?TTTTAGGT
GATCCCAA?ATCATGCG?TCGATCCA?TGGATGAC?GTTTGGAA
TGCGACCC?CAGGATGG?GCGAGGTC?ACCCGCTG?AGTTTAAG
CATATCAA?TAAGCGGA?GGAGAAGA?AACTTACG?AGGATTCC
CCTAGTAA?CGGCGAGT
22. the ITS2 of the poly-stem of noble dendrobium (Dendrobium lindleyi Stendel) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AACCATCG?AGTCTTTG?AACGCAAG?TTGCGCCC?GAGGCCAA
TCGGCCAG?GGCACGTT?CGCCTGGG?CGTCAAGC?ATTATGAC
GCTCCCTG?TCAACTCC?ATCCCATC?TTTTAATG?GGCTGGCG
AAGGCTCG?GATGTGCA?AAGTGGCT?CGTCGTGC?CTCTCAGT
GCGGCGGG?CTTAAGAG?CGGGTCAT?CATCTCGT?TGGCTACG
AACAATTA?CGGGTGGA?TTAAAGTG?AGGCCTAT?ATTATATT
GTCGTGTA?TGCCTGAG?AGAAGATT?ATTCATTT?TTTGGTTA
TCCCAAAT?CATGCGTC?GATCCATA?GATCACGT?TTGGAATG
CGACCCCA?GGATGGGC?GAGGTCAC?CCGCTGAG?TTTAAGCA
TATCACTA?AGCGGAGG?AGAAGAAA?CTTACGAG?GATTCCCC
TAGTAACG?GCGAGT
23. the ITS2 of the poly-stem of noble dendrobium (Dendrobium lindleyi Stendel) sample 2 and 5.8S and 28S be the rDNA sequence partly:
AACCATCG?AGTCTTTG?AACGCAAG?TTGCGCCC?GAGGCCAA
TCGGCCAA?GGGCACGT?TCGCCTGG?GCGTCAAG?CATTATGA
CGCTCCCT?GTCAACTC?CATCCCAT?CTTTTAAT?GGGCTGGC
GAAGGCTC?GGATGTGC?AAAGTGGC?TCGTCGTG?CCTCTCAG
TGCGGCGG?GCTTAAGA?GCGGGTCA?TCATCTCG?TTGGCTAC
GAACAATT?ACGGGTGG?ATTAAAGT?GAGGCCTA?TATTATAT
TGTCGTGT?ATGCCTGA?GAGAAGAT?TATTCATT?TTTTGGTT
ATCCCAAA?TCATGCGT?CGATCCAT?AGATCACG?TTTGGAAT
GCGACCCC?AGGATGGG?CGAGGTCA?CCCGCTGA?GTTTAAGC
ATATCACT?AAGCGGAG?GAGAAGAA?ACTTACGA?GGATTCCC
CTAGTAAC?GGCGAGT
24. the ITS2 of the U.S. flower stem of noble dendrobium (Dendrobium loddigesii Rolfe) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCG?AGG
CCAACCGG?CCAAGGGC?ACGTCCGC?CTGGGCGT?CAGTCATT
TTATCGCT?CTGTGCCA?AGCTACCC?ATCGATTG?ATGTGTTG
GCGAAGGC?TTGGATGT?GCATTATG?GCTCATCG?TGCCCCTG
GGCGCGGC?GGGCTGAA?GAGCGGGT?CATCATCT?CGTTGGCT
GCGAACAA?TAAGGGGT?GGACTGAA?GGGAGACC?TATGCTAT
TGTGTCGT?GCATGCCT?AAGAGTTG?ATCATACC?TTTGAGGA
GATCCCAA?ATCATGCG?TCGATCTA?TGGACGGC?GCTTTGAA
TGCGACCC?CAGGATGG?GCGAGGCC?ACCCGCCG?AGTTTAAG
CATATCAA?TAAGCGGA?GGAGAAGA?AACTTACG?AGGATTCC
CCTAGTAA?CGGCGAGT
25. the ITS2 of the U.S. flower stem of noble dendrobium (Dendrobium loddigesii Rolfe) sample 2 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTT?GCGC?CCGAG?GCC
AACCGGCC?AAGGGCAC?GTCCGCCT?GGGCGTCA?GTCATTTT
ATCGCTCT?GTGCCAAG?CTAACCAT?CGATTGAT?GTGTTGGC
GAAGGCTT?GGATGTGC?ATTATGGC?TCATCGTG?CCCCTGGG
CGCGGCGG?GCTGAAGA?GCGGGTCA?TCATCTCG?TTGGCTGC
GAACAATA?AGGGGTGG?ACTGAAGG?GAGACCTA?TGCTATTG
TGTCGTGC?ATGCCTAA?GAGTTGAT?CATACCTT?TGAGGAGA
TCCCAAAT?CATGCGTC?GATCTATG?GACGGCGC?TTTGAATG
CGACCCCA?GGATGGGC?GAGGCCA?CCCGCCGA?GTTTAAGC
ATATCAAT?AAGCGGAG?GAGAAGAA?ACTTACGA?GGATTCCC
CTAGTAAC?GGCGAGT
26. the ITS2 of the U.S. flower stem of noble dendrobium (Dendrobium loddigesii Rolfe) sample 3 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCGAGGCC
AACCGGCC?AAGGGCAC?GTCCGCCT?GGGCGTCA?GTCATTTT
ATCGCTCT?GTGCCAAG?CTACCCAT?CGATTGAT?GTGTTGGC
GAAGGCTT?GGATGTGC?ATTATGGC?TCATCGTG?CCCCTGGG
CGCGGCGG?GCTGAAGA?GCGGGTCA?TCATCTCG?TTGGCTGC
GAACAATA?AGGGGTGG?ACTGAAGG?GAGACCTA?TGCTATTG
TGTCGTGC?ATGCCTAA?GAGTTGAT?CATACCTT?TGAGGAGA
TCCCAAAT?CATGCGTC?GATCTATG?GACGGCGC?TTTGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CCGCCGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AGAAGAAA?CTTACGAG?GATTCCCC
TAGTAACG?GCGAGT
27. the ITS2 of sieve river stem of noble dendrobium (Dendrobium lohohense T.Tang et F.T.Wang) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AACCATCG?AGTCTTTG?AACGCAAG?TTGCGCCC?GAGGCCAA
CCGGCCAA?GGGCACGT?CTGCCTGG?GCGTCAAG?CATTACAT
CGCTCCAT?GTCAAGTC?ACCCTTCG?ATGGATGG?GCTGGCGA
AGGCTTGG?ATGTGCAC?AGTGGCTC?GTCGTGCC?ACTTGGTG
CGGCGGGC?TGAAGAGC?GGGTCGTC?ATCCTCTT?GGCCGCGA
ACAATAAT?GGGTGGAT?TAAAGCGA?TGCCTAAG?TTATTGTG
TGTGCATG?CCTAAGAG?ATGATTAT?ACTTTTTA?AGTGATCC
CAATTCAT?GCGTCGAT?CCATGGAT?GACGTTTT?GAATGCGA
CCCCAGGA?TAGGCGAG?GCCACCCG?CCGAGTTT?AAGCATAT
CAATAAGC?GGAGGAGA?AGAAACTT?ACGAGGAT?TCCCCTAG
TAACGGCG?AGT
28. the ITS2 of dendrobium moniliformeSweet (Dendrobium moniliforme (L.) Sw.) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CTGAGGCC
AACCGGCT?GAGGGCAC?GTCCGCCT?GGGCGTCA?AGCATTTT
ATCACTCC?GTGCCTAC?TCTCCCAT?CCATGGAT?GTGTTGCT
AAGGCTCG?GATGTGCA?CGGTGGCT?CGTCGTGC?CCCTTGGT
GCGGCGGG?CTGAAGGG?CGGGTCAT?CTTCTCGT?TGGCTGCC
AACAATAA?GGGGTGGA?TTAAATAA?GGCCTATG?CTATTGTG
TCAAGCGC?GCCCGAGA?GATGGTCA?TACTTTTT?AGGTGATC
CCAATTCA?TGCGTTGA?TCCATGGA?TGGCGTAT?CGAATGTG
ACCCCAGG?ATGGGCGA?GGCCACCC?GCTGAGTT?TAAGCATA
TCAATAAG?CGGAGGAG?AAGAAACT?TACGAGGA?TTCCCCTA
GTAACGGC?GAGT
29. the ITS2 of dendrobium moniliformeSweet (Dendrobium moniliforme (L.) Sw.) sample 2 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CTGAGGCC
AACCGGCT?GAGGGCAC?GTCCGCCT?GGGCGTCA?AGCATTTT
ATCACTCC?GTGCCTAC?TCTCCCAT?CCATGGAT?GTGTTGCT
AAGGCTCG?GATGTGCA?CGGTGGCT?CGTCGTGC?CCCTTGGT
GCGGCGGG?CTGAAGGG?CGGGTCAT?CTTCTCGT?TGGCTGCC
AACAATAA?GGGGTGGA?TTAAATAA?GGCCTATG?CTATTGTG
TCAAGCGC?GCCCGAGA?GATGGTCA?TACTTTTT?AGGTGATC
CCAATTCA?TGCGTTGA?TCCATGGA?TGGCGTAT?CGAATGTG
ACCCCAGG?ATGGGCGA?GGCCACCC?GCTGAGTT?TAAGCATA
TCAATAAG?CGGAGGAG?AAGAAACT?TACGAGGA?TTCCCCTA
GTAACGGC?GAGT
30. the ITS2 of dendrobium moniliformeSweet (Dendrobium moniliforme (L.) Sw.) sample 3 and 5.8S and 28S be the rDNA sequence partly:
AACCATCG?AGTCTTTG?AACGCAAG?TTGCGCCT?GAGGCCAA
CCGGCTGA?GGGCACGT?CCGCCTGG?GCGTCAAG?CATTTTAT
CACTCCGT?GCCTACTC?TCCCATCC?ATGGATGT?GTTGCTAA
GGCTCGGA?TGTGCACG?GTGGCTCG?TCGTGCCC?CTTGGTGC
GGCGGGCT?GAAGGGCG?GGTCATCT?TCTCGTTG?GCTGCCAA
CAATAAGG?GGTGGATT?AAATAAGG?CCTATGCT?ATTGTGTC
AAGCGCGC?CCGAGAGA?TGGTCATA?CTTTTTAG?GTGATCCC
AATTCATG?CGTTGATC?CATGGATG?GCGTATCG?AATGTGAC
CCCAGGAT?GGGCGAGG?CCACCCGC?TGAGTTTA?AGCATATC
AATAAGCG?GAGGAGAA?GAAACTTA?CGAGGATT?CCCCTAGT
AACGGCGA?GT
31. the ITS2 of the spoon lip stem of noble dendrobium (Dendrobium moschatum (Buch.-Ham.) Sw.) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CTG?AGG
CCAACCGG?CCAAGGGC?ACGTCTGC?CTGGGCGT?CAAGCGAT
ATGTCGCT?CTGTGCTA?AGTTAACC?ATCGATGG?ATGGGTTG
GCGAAGGC?TCGGATGT?GCATAATG?GCTCGTTG?TGCCCCTC
AGCGTGGC?GGGCTGAA?GAGGGGGT?CATCATCT?TGTTAGTC
GCAAACAA?TAAGGGGT?GGATAAAT?ATGAGGCC?TATGTTAT
TGTGTTGT?GCATGCCC?AAGAGATG?ATTAAACC?TTTTTTGT
GATCCTAA?ATCATGCG?TCAATCCA?TGTATGGC?GTTTTGAA
TGCGACCC?CAGGATGG?GCGAGGCC?ACCCGCCG?AGTTTAAG
CATATCAA?TAAGCGGA?GGAGAAGA?AACTTACA?AGGATTCC
CCTAGTAA?CGGCGAGT
32. the ITS2 of the spoon lip stem of noble dendrobium (Dendrobium moschatum (Buch.-Ham.) Sw.) sample 2 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CTGAGGCC
AACCGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGCGATAT
GTCGCTCT?GTGTTAAG?TTAACCAT?CGATGGAT?GGGTTGGC
GAAGGCTC?GGATGTGC?ATAATGGC?TCGTTGTG?CCCCTCAG
CGTGGCGG?GCTGAAGA?GGGGGTCA?TCATCTTG?TTAGTCGC
AAACAATA?AGGGGTGG?ATAAATAT?GAGGCCTA?TGTTATTG
TGTTGTGC?ATGCCCAA?GAGATGAT?TAAACCTT?TTTTGTGA
TCCTAAAT?CATGCGTC?AATCCATG?TATGGCGT?TTTGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CCGCCGAG?TTTAAGCA
TATCAATA?AGCGGTGG?AGAAGAAA?CTTACAAG?GATTCCCC
TAGTAACG?GCGAGT
33. the ITS2 of the spoon lip stem of noble dendrobium (Dendrobium moschatum (Buch.-Ham.) Sw.) sample 3 and 5.8S and 28S be the rDNA sequence partly:
AACCATCG?AGTCTTTG?AACGCAAG?TTGCGCCT?GAGGCCAA
CCGGCCAA?GGGCACGT?CTGCCTGG?GCGTCAAG?CGATATGT
CGCTCTGT?GCTAAGTT?AACCATCG?ATGGATGG?GTTGGCGA
AGGCTCGG?ATGTGCAT?AATGGCTC?GTTGTGCC?CCTCAGCG
TGGCGGGC?TGAAGAGG?GGGTCATC?ATCTTGTT?AGTCGCAA
ACAATAAG?GGGTGGAT?AAATATGA?GGCCTATG?TTATTGTG
TTGTGCAT?GCCCAAGA?GATGATTA?AACCTTTT?TTGTGATC
CTAAATCA?TGCGTCAA?TCCATGTA?TGGCGTTT?TGAATGCG
ACCCCAGG?ATGGGCGA?GGCCACCC?GCCGAGTT?TAAGCATA
TCAATAAG?CGGAGGAG?AAGAAACT?TACAAGGA?TTCCCCTA
GTAACGGC?GAGT
34. the ITS2 of dendrobium stem (Dendrobium nobile Lindl.) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GA?ACGCAAGT?TGCGCCTG?AGGC
CAATCGGT?TGAGGGCA?CGTCCGCC?TGGGCGTC?AAGCAT
TTTATCGC?TCCGTGCC?TAGTCTCCCA?TCCATGGA?TGTGTTGC
CAAGGCTC?GGATGTGC?ACGGTGGC?TCGTCGTG?CCCGTTGG
TGCGGCGG?GCTGAAGG?GCGGGTCA?TCTTCTCG?TTGGTTGC
CAACAATA?AGGGGTGG?ATTAAATA?AGGCCTAT?GCTATTGT
GTCAAGCG?CGCCTGAG?AGATGGTC?ATACTTTT?TAGGTGAT
CCCAATTC?ATGCGTCG?ATCCATGG?ATGGCGTA?TCGAATGT
GACCCCAG?GATGGGCG?AGGCCACC?CGCCGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?GAAGAAAC?TTACGAGG?ATTCCCCT
AGTAACGG?CGAGT
35. the ITS2 of dendrobium stem (Dendrobium nobile Lindl.) sample 2 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CTG?AGG
CCAACCGG?CTGAGGGC?ACGTCCGC?CTGGGCGT?CAAGCATT
TTATCGCT?CCGTGCCT?AGTCTCCC?ATCCATGG?ATGTGTTG
CCAAGGCT?CGGATGTG?CACGGTGG?CTCGTCGT?GCCCCTTG
GTGCGGCG?GGCTGAAG?GGCGGGTC?ATCTTCTC?GTTGGCTG
CCAACAAT?AAGGGGTG?GATTAAAT?AAGGCCTA?TGCTATTG
TGTCAAGC?GCGCCTGA?GAGATGGT?CATACTTA?TTAGGTGA
TCCCAATT?CATGCGTC?GATCCATG?GATGGCGT?ATCGAATG
TGACCCCA?GGATGGGC?GAGGCCAC?CCGCCGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AGAAGAAA?CTTACGAG?GATTCCCC
TAGTAACG?GCGAGT
36. the ITS2 of dendrobium stem (Dendrobium nobile Lindl.) sample 3 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CTGAGG
CCAACCGG?CTGAGGGC?ACGTCCGC?CTGGGCGT?CAAGCATT
TTATCGCT?CCGTGCCT?AGTCTCCC?ATCCATGG?ATGTGTTG
CCAAGGCT?CGGATGTG?CACGGTGG?CTCGTCGT?GCCCCTTGG
TGCGGCGG?GCTGAAGG?GCGGGTCA?TCTTCTCG?TTGGCTGC
CAACAATA?AGGGGTGG?ATTAAATA?AGGCCTAT?GCTATTGT
GTCAAGCG?CGCCTGAG?AGATGGTC?ATACTTAT?TAGGTGAT
CCCAATTC?ATGCGTCG?ATCCATGG?ATGGCGTA?TCGAATGT
GACCCCAG?GATGGGCG?AGGCCACC?CGCCGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?GAAGAAAC?TTACGAGG?ATTCCCCT
AGTAACGG?CGAGT
37. the ITS2 of Herba Dendrobii (Dendrobium officinale Kimura et Migo) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCA?AGG
CCAACCGG?CTAAGGGC?ACGTCCGC?CTGGGCGT?CAAGCATT
TTATCTCT?CCGTGCCT?AATCTCCC?ATCCATGG?ATGTGTTA
CTAAGGCT?CGGATGTG?CATGGTGG?CTCCTCGT?GCCCCTTG
GTGCGGCG?GGCTGAAG?GGCGGGTC?ATCTTCTC?GTTGGTTG
CCAACAAT?AAGGGGTG?GATTAAAA?AAGGCCTA?TGCTATTG
TGATAAGC?GCGCCCGA?GAGATGAT?CATACTTT?TTAGGTGA
TCCCAATC?CATGCGCT?AATCCATG?GATGGCGT?ATCGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CTGCTGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AAAAGAAA?CTTACGAG?GATTCCCC
TAGTAACG?GCGAGT
38. the ITS2 of Herba Dendrobii (Dendrobium officinale Kimura et Migo) sample 2 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCAAGGCC
AACCGGCT?AAGGGCAC?GTCCGCCT?GGGCGTCA?AGCATTTT
ATCTCTCC?GTGCCTAA?TCTCCCAT?CCATGGAT?GTGTTACT
AAGGCTCG?GATGTGCA?TGGTGGCT?CCTCGTGC?CCCTTGGT
GCGGCGGG?CTGAAGGG?CGGGTCAT?CTTCTCGT?TGGTTGCC
AACAATAA?GGGGTGGA?TTAAAA?A?GGCCTATG?CTATTGTG
ATAAGCGC?GCCCGAGA?GATGATCA?TACTTTTT?AGGTGATC
CCAATCCA?TGCGCTAA?TCCATGGA?TGGCGTAT?CGAATGCG
ACCCCAGG?ATGGGCGA?GGCCACCT?GCTGAGTT?TAAGCATA
TCAATAAG?CGGAGGAA?AAGAAACT?TACGAGGA?TTCCCCTA
GTAACGGC?GAGT
39. the ITS2 of Herba Dendrobii (Dendrobium officinale Kimura et Migo) sample 3 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCAAGGCC
AACCGGCT?AAGGGCAC?GTCCGCCT?GGGCGTCA?AGCATTTT
ATCTCTCC?GTGCCTAA?TCTCCCAT?CCATGGAT?GTGTTACT
AAGGCTCG?GATGTGCA?TGGTGGCT?CCTCGTGC?CCCTTGGT
GCGGCGGG?CTGAAGGG?CGGGTCAT?CTTCTCGT?TGGTTGCC
AACAATAA?GGGGTGGA?TTAAAAAA?GGCCTATG?CTATTGTG
ATAAGCGC?GCCCGAGA?GATGATCA?TACTTTTT?AGGTGATC
CCAATCCA?TGCGCTAA?TCCATGGA?TGGCGTAT?CGAATGCG
ACCCCAGG?ATGGGCGA?GGCCACCT?GCTGAGTT?TAAGCATA
TCAATAAG?CGGAGGAA?AAGAAACT?TACGAGGA?TTCCCCTA
GTAACGGC?GAGT
40. the ITS2 of Herba Dendrobii (Dendrobium officinale Kimura et Migo) sample 4 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCAAGGCC
AACCGGCT?AAGGGCAC?GTCCGCCT?GGGCGTCA?AGCATTTT
ATCTCTCC?GTGCCTAA?TCTCCCAT?CCATGGAT?GTGTTACT
AAGGCTCG?GATGTGCA?TGGTGGCT?CCTCGTGC?CCCTTGGT
GCGGCGGG?CTGAAGGG?CGGGTCAT?CTTCTCGT?TGGTTGCC
AACAATAA?GGGGTGGA?TTAAAAAA?GGCCTATG?CTATTGTG
ATAAGCGC?GCCCGAGA?GATGATCA?TACTTTTT?AGGTGATC
CCAATCCA?TGCGCTAA?TCCATGGA?TGGCGTAT?CGAATGCG
ACCCCAGG?ATGGGCGA?GGCCACCT?GCTGAGTT?TAAGCATA
TCAATAAG?CGGAGGAA?AAGAAACT?TACGAGGA?TTCCCCTA
GTAACGGC?GAGT
41. the ITS2 of Herba Dendrobii (Dendrobium officinale Kimura et Migo) sample 5 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCAAGGCC
AACCGGCT?AAGGGCAC?GTCCGCCT?GGGCGTCA?AGCATTTT
ATCTCTCC?GTGCCTAA?TCTCCCAT?CCATGGAT?GTGTTACT
AAGGCTCG?GATGTGCA?TGGTGGCT?CCTCGTGC?CCCTTGGT
GCGGCGGG?CTGAAGGG?CGGGTCAT?CTTCTCGT?TGGTTGCC
AACAATAA?GGGGTGGA?TTAAAAAA?GGCCTATG?CTATTGTG
ATAAGCGC?GCCCGAGA?GATGATCA?TACTTTTT?AGGTGATC
CCAATCCA?TGCGCTAA?TCCATGGA?TGGCGTAT?CGAATGCG
ACCCCAGG?ATGGGCGA?GGCCACCT?GCTGAGTT?TAAGCATA
TCAATAAG?CGGAGGAA?AAGAAACT?TACGAGGA?TTCCCCTA
GTAACGGC?GAGT
42. the ITS2 of Herba Dendrobii (Dendrobium officinale Kimura et Migo) sample 6 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCAAGGCC
AACCGGCT?AAGGGCAC?GTCCGCCT?GGGCGTCA?AGCATTTT
ATCTCTCC?GTGCCTAA?TCTCCCAT?CCATGGAT?GTGTTACT
AAGGCTCG?GATGTGCA?TGGTGGCT?CCTCGTGC?CCCTTGGT
GCGGCGGG?CTGAAGGG?CGGGTCAT?CTTCTCGT?TGGTTGCC
AACAATAA?GGGGTGGA?TTAAAAAA?GGCCTATG?CTATTGTG
ATAAGCGC?GCCCGAGA?GATGATCA?TACTTTTT?AGGTGATC
CCAATCCA?TGCGCTAAT?CCATGGAT?GGCGTATC?GAATGCGA
CCCCAGGA?TGGGCGAG?GCCACCTG?CTGAGTTT?AAGCATAT
CAATAAGC?GGAGGAAA?AGAAACTT?ACGAGGAT?TCCCCTAG
TAACGGCG?AGT
43. the ITS2 of Dendrobium williamsonii Day et Reichb. F. (Dendrobium williamsonii Day et Rchb.f.) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AA?GTT?GCG?CCT?GAG?GCC
AACCGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGCGTTAC
GTCACTCC?GTGCCAAC?TCCATTCC?ATCGATGG?ATGGGCCG
GTGAAGGC?TCGGATGT?GTAGAGTG?GCTCGTCG?TGCCCGTC
GGTGCGGC?GGGTTGAA?GAGTGGGT?CATCATCTC?GTTGGCTG
CGAACAAT?AAGGGGTG?GAATAAAG?TGAGACCT?ATGTTATT
GTGTCGTG?TATGCCTG?AGAGAAGA?CCATTATC?AGGAGATC
CCAAATCA?TGCGTCGA?TCCATGAA?TGGCGTTT?GGAATGCG
ACCCCAGG?ATGGGCGA?GGCCACCC?GCCGAGTT?TAAGCATA
TCAATAAG?CGGAGGAG?AAGAAACT?TACGAGGA?TTCCCCTA
GTAACGGC?GAGT
44. the ITS2 of Dendrobium williamsonii Day et Reichb. F. (Dendrobium williamsonii Day et Rchb.f.) sample 2 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CTGAGG
CCAACCGG?CCAAGGGC?ACGTCTGC?CTGGGCGT?CAAGCGTT
ACGTCACT?CCGTGCCA?ACTCCATT?CCATCGAT?GGATGGGC
CGGTGAAG?GCTCGGAT?GTGTAGAG?TGGCTCGT?CGTGCCCG
TCGGTGCG?GCGGGTTG?AAGAGTGG?GTCATCAT?CTCGTTGG
CTGCGAAC?AATAAGGG?GTGGAATA?AAGTGAGA?CCTATGTT
ATTGTGTC?GTGTATGC?CTGAGAGA?AGACCATT?ATCAGGAG
ATCCCAAA?TCATGCGT?CGATCCAT?GAATGGCG?TTTGGAAT
GCGACCCC?AGGATGGG?CGAGGCCA?CCCGCCGA?GTTTAAGC
ATATCAAT?AAGCGGAG?GAGAAGAA?ACTTACGA?GGATTCCC
CTAGTAAC?GGCGAGT
45. the ITS2 of Pholidota cantonensis (Pholidota cantonensis Rolfe) sample 1 and 5.8S and 28S be the rDNA sequence partly:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCGAGGCC
AACCGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGCGTTGC
GTCGCTCC?GTGCCATC?CCCGTCCC?ACGAATGC?GTGCGACG
GTCGAGGC?CCGGACGT?GCAGAGTG?GCTCGTCG?TGCCCGTT
GGTGCGGC?GGGCTGAA?GAGTGTGG?GCTATCGT?CTCGTTGG
CCAACAGC?GACAAGGG?GTGGATGA?AAGCTGTA?TGAGCAAA
GAGAGCCT?ATGTTGTT?GCTTGCTT?GGCCCGAG?AAAAGATT
CCCAGCTG?ATGATCCC?AACCCACG?CGTCGATC?CACAGGCG
ACGGCTTG?GAATGCGA?CCCCAGGA?TGGGCGAG?GCCACCCG
CCGAGTTT?AAGCATAT?CAATAAGC?GGAGGAGA?AGAAACTT
ACGAGGAT?TCCCCTAG?TAACGGCG?AGT
46. the ITS2 of Pholidota cantonensis (Pholidota cantonensis Rolfe) sample 2 and 5.8S and 28S be the rDNA sequence partly:
AACCATCG?AGTCTTTG?AACGCAAG?TTGCGCCC?GAGGCCAA
CCGGCCAA?GGGCACGT?CTGCCTGG?GCGTCAAG?CGTTGCGT
CGCTCCGT?GCCATCCC?CGTCCCAC?GAATGCGT?GCGACGGT
CGAGGCCC?GGACGTGC?AGAGTGGC?TCGTCGTG?CCCGTTGG
TGCGGCGG?GCTGAAGA?GTGTGGGC?TATCGTCT?CGTTGGCC
AACAGCGA?CAAGGGGT?GGATGAAA?GCTGTATG?AGCAAAGA
GAGCCTAT?GTTGTTGC?TTGCTTGG?CCCGAGAA?AAGATTCC
CAGCTGAT?GATCCCAA?CCCACGCG?TCGATCCA?CAGGCGAC
GGCTTGGA?ATGCGACC?CCAGGATG?GGCGAGGC?CACCCGCC
GAGTTTAA?GCATATCA?ATAAGCGG?AGGAGAAG?AAACTTAC
GAGGATTC?CCCTAGTA?ACGGCGAGT
The measuring method of the ITS2 of above-mentioned 46 samples and 5.8S and 28S rrna partial dna sequence comprises the following steps:
(1) collects plant sample, carry out cultivar identification work;
(2) from the plant that step (1) is selected, extract DNA;
(3) DNA that extracts with step (2) is a template, with the primer of oligonucleotide as polymerase chain reaction (PCR), and the section of amplification ITS2 and part 5.8S and 28S, used primer divides Do to pick up from the conservative section of plant 5.8S and 28S;
(4) using direct sequencing technologies checks order to the PCR product that is obtained in the step (3).
Each sample repeats to check order 4 times at least.And the sequence of each sample also internet page National Center for Biotechnology Information (NCBI) and GeneticsComputer Group package (GCG) make comparisons with the sequence of other cymbidium varieties and verify to guarantee 46 sequences of the present invention accurate.
As implied above, except the stem of noble dendrobium of sieve river, the hundred parts of rates of variation at dna level between the sample of measured same kind are not higher than 1.22%.The difference of dna sequence dna between the kind of described 16 kinds of stems of noble dendrobium (ITS2 and part 5.8S and 28S) is 2% to 17%, and the difference of the dna sequence dna of the stem of noble dendrobium and Pholidota cantonensis (ITS 2 and part 5.8S and 28S) is 16% to 20%.
The present invention utilizes ITS2 (transcribing interstice coverage 2) in the stem of noble dendrobium class plant and between partly the dna sequence dna of 5.8S and 28S plant and plant interior comparison, identifies different stem of noble dendrobium kinds whereby and distinguish its true and false.The method that is adopted comprises the following steps:
(1) from plant, extracts DNA;
(2) be template with the DNA that extracts in the step (1), with the primer of oligonucleotide as polymerase chain reaction (PCR), the section of amplification ITS2 and part 5.8S and 28S, used primer divides Do to pick up from the conservative section of plant 5.8S and 28S;
(3) using direct sequencing technologies checks order to the PCR product that is obtained in the step (2);
(4) with the plant ITS2 that records in the step (3) and partly the dna sequence dna comparison of dna sequence dna and above-mentioned 46 stems of noble dendrobium and the Pseudobulbus Pholidotae Chinensis sample of 5.8S and 28S with the stem of noble dendrobium kind of evaluation institute measuring plants or offer an explanation its true and false.
Describe described method below in detail:
(1) extraction of DNA
DNA of plants extracting method and Draper and Scott (Draper and Scott, 1988) are described roughly the same, are summarized as follows: the exsiccant vegetable material is washed with 70% alcoholic acid distilled water solution, then, mix and grind into powder with liquid nitrogen.The powdered sample of gained and the CTAB of 12 times of volumes are mixed Uniform, and insulation added chloroform-primary isoamyl alcohol (24: 1) mixed solution of equivalent after 30 minutes in 56 ℃ of water-baths, and is centrifugal, gets supernatant liquor.The 10%CTAB solution that adds 0.1 volume in resistates, room temperature is placed after 10 minutes and extracted once with chloroform-primary isoamyl alcohol, and is centrifugal, gets supernatant liquor and mixes with last time supernatant liquor.The CTAB that adds equivalent in the gained supernatant liquor produces precipitation, and room temperature was placed after 1 hour with 13000g centrifugal 15 minutes.Be dissolved in the TE damping fluid (10mM Tris-HCl, pH 7.5,1mM EDTA) after with ethanol purification the DNA throw out standby.
(2) pcr amplification reaction
The DNA of ITS2 and part 5.8S and 28S section is obtained by pcr amplification reaction.The primer that reacts used is 5.8d and 28cc (Hillis and Dixon, 1991):
5.8d pick up from the conservative section of plant 5.8S
5’AAC?CAT?CGA?GTC?TTT?GAA?CGC?A?3’
28cc picks up from the conservative section of plant 28S
5’ACT?CGC?CGT?TAC?TAG?GGG?AA?3’
Reaction soln comprises: 1ng DNA of plants, 1Xtaq damping fluid (10mM Tris-HCl, pH8.3,0.001% gelatin), 0.2mM dNTPs, 1 μ M primer (5.8d and 28cc), and the Taq DNA polymerase of 1 unit.Reaction conditions is: in 94 ℃ of pre-treatment 5 minutes, carry out 35 circulations then, cycling condition is: 94 ℃, and 1 minute, 60 ℃, 1 minute, 72 ℃, 2 minutes; Last 72 ℃, 10 minutes.
(3) direct sequencing technologies
The PCR product that step (2) is obtained with the ethanol alcoholization after, take out 1 μ L (about 50ng) PCR and refine product, with 4 μ L reaction terminating mixed solutions, 0.64 μ L, 25 μ M primers (5.8d and 28cc), and the mixing of 4.86 μ L distilled water.Carry out following 25 circulations then, reaction conditions is: 96 ℃, and 10 seconds, 56 ℃, 5 seconds, 60 ℃, 4 minutes.Use sequence analysis software (" the Sequencing Analysis third edition " software and Hitachi " DNASIS the 7th edition ") analysis of Perkin Elmer and the dna sequence dna of definite sample at last.
Therefore, adopt the ITS2 of the stem of noble dendrobium provided by the invention and the partly dna sequence dna and the authentication method of 5.8S and 28S section,, make convenient and reliable the evaluation of the valuable Chinese medicine stem of noble dendrobium by its ITS2 and the partly comparison of the dna sequence dna of 5.8S and 28S section.

Claims (3)

1. the ITS2 of a stem of noble dendrobium and 5.8S and 28S rrna partial dna sequence comprise:
(1) ITS2 of hooked dendrobium and 5.8S and 28S rrna part rDNA sequence:
AAC?CATCGA?GTCTTT?GAA?CGC?AAGTTGCGC?CCGAGGCCAA
CCGGCTAA?GGGCACGT?CCGCCTGG?GCGTAAGC?ATTTTATC
GCTCCGTG?CCTAATCT?CCCATCCA?TGTATGTG?TTGCAAAG
GCTCGGAT?GTGCACGG?TGGCTCCT?CGTGCCCC?TTGGTGCG
GCGGGCTG?AAGGCCGG?GTCATCTT?CTCGTTGG?TTGCCAAC
AATAAGGG?GTGGATTA?AACAAGGC?CTATGCTG?TTGTGATA
TGCGCGCC?CGAGAGAT?GATCATAC?TTTTTAGG?TGATCCCA
ATTCATGC?GCTGATCC?ATGGATGG?CGTATCGA?ATGCGACC
CCAGGATG?GGCGAGGC?CACCCGCT?GAGTTTAA?GCATATCA
GCATATCAAT?AAG?CGGAGG?AAAA?GAAA?CTTACGAGGATT
CCC?CTA?GTA?ACG?GCG?AGT；
(2) ITS2 of Dendrobium chrysanthum and 5.8S and 28S rrna part rDNA sequence:
AACCATCGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCCGAGGC
CAACCTGC?CAAGGGCA?CGTCTGCC?TGGGCGTC?AAGCATTT
TGTTGCTC?CGTGTCTC?GTCACCCA?TCAATGGA?TGTGCTGG
CGAAGGCT?CGGATGTG?CACGGTGG?CCCGTCGT?GCCCCTCG
GTGCGGCG?GGCTTAAG?AGCGGGTC?GCCATCTC?GTTGGTTG
CGAACAAT?AAGGGGTG?GATTAAAT?GGAGGCCT?ATGCTATT
GTGTCGTG?CATGCCTA?AGAGATTA?AATTTTTT?AGGTGATC
CCAAATCA?CGCGTCGA?TCCACGGA?TGATGCTT?TGAATGCG
ACCCCAGG?ATGGGCGA?GGCCACCC?GCCGAGTT?TAAGCATA
TCAATAAG?CGGAGGAG?AAGAAACT?TACAAGGA?TT?CCC
CTA?GTA?ACG?GCG?AGT；
(3) ITS2 of Dendrobium chrysotoxum and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCCGAGGC
CAATCGGC?CAAGGGCA?CGTCTGCC?TGGGCGTC?AAGCGTTA
CGTCGCTC?CGTGTCCA?GCCACCCG?TCGATGGA?TGGGCTGG
CGATGGCT?CGGATGTG?CACAGTGG?CTCCTCGT?GCCCCTCG
GCACGGCG?GGCTTAAG?AGCGGGTA?ATCATCTC?GTTGGCTG
CGAACAAT?ATGGGGTG?GATAAATG?CGATGCTT?ATGATATT
GTGCCGTG?CATGCCCG?AGAGATGA?TCATACTT?ATTCAGGT
GATCCCAA?ACCATGCG?TCGATCCA?TGGATGGC?GTTTTGAA
TGCGACCC?CAGGACAG?GCGAGGCC?ACCCGCCG?AGTTTAAG
CATATCAA?TAAGCGGA?GGAGAAGA?AACTTACG?AGGATTCC
CCTAGTAA?CGGCG?AGT
(4) ITS2 of Dendrobium crepidatum lindl et paxt and 5.8S and 28S rrna part rDNA sequence:
AACCAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCG?AGG?CCA
ATCGGCCA?AGGGCACG?TCTGCCTG?GGCGTCAA?GCATTTTG
TTGCTCCG?TGTCTACT?CACACCCA?TCGATGGA?TGTGCTGG
TGTAGGCT?CGGATGTG?CATGCTGG?CTCGTCGT?TCTCCCAG
GTGCGGCG?GGCTGAAG?AGCGGGTC?ATCCTCTC?GTTTGCGG
CGAACAAT?AAGGGGTG?GATTGAAG?AGAGGTCT?ATGCTATT
GTGTCGTG?CATGCCTG?AGAGATGA?TTAACTTT?TTTGTGAT
CCTAAATC?ATGCATCG?ATCCATGG?ATGATGCT?TTGAATGT
GACCCCAG?GATGGGCG?AGGCTACC?CGCTGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?GAAGAAAC?TTACGAGG?A?TT?CCC
CTA?GTA?ACG?GCG?AGT
(5) ITS2 of dendrobium and 5.8S and 28S rrna part rDNA sequence:
AACCAT?CGA?GTC?TTT?GAA?CGC?A?AG?TTGCGCCCGAGG
CCAACCGG?CCAAGGGC?ACGTCTGC?CTGGGCGT?CAAGCGTT
ATGTCGCT?CCGTGTTA?ACTCCATC?CCATCGAT?GTATGGGC
TGGCGAAG?GCTCGGAT?GTGCAGTG?TGGCTCGT?CGTGCCCC
TTGGTGCG?GCGAGCTG?AAAAGCGG?GTAATCAT?CTCGTTGG
CTGCGAAC?GATAAGGG?GTGGATTA?AAGCGAGA?CCTATGTT
ATTGTGTC?GTGTATGC?CCGAGAGA?AGATTATA?CATACTCA
GGTGATCC?TGAATCAT?GCGTCGAT?CAAAGGAC?GGCGCTTG
GAATGCGA?CCCCAGGA?TGGGCGAG?GCCACCCG?CCGAGTTT
AAGCATATT?AATAAGCG?GAGGAGAA?GAAACTTA?CGAGGATT
CCCCTAGT?A?ACG?GCG?AGT
(6) ITS2 of Dendrobium fimbriatum Hook. and 5.8S and 28S rrna part rDNA sequence:
AAC?CATCGAGTC?TTTGAACGCA?AGTTGCGC?CCGAGGCC
AACAGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGTGTTAT
GTCGCTCC?ATGCCAAG?TCACCCAT?CGATGGAT?GGGCCGGC
GAAGGCTC?GGATGTGC?ACAGTGGC?TCGCCGTG?CCCCTCGG
TGCGGCGG?GCTGAAGA?GCGGGTCA?TTATCTTG?TTGGCTGC
AGGCAATA?ATGGGTGG?ATAAACGC?GAGGCCTT?TGTTATTG
TGTCGTGC?ATGCCTAA?GATATGAT?TAAACTTT?TTAGGTGA
TCCCAAAT?CATGCGTC?GATCCACG?GATGGCGT?TTTGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CCGCCGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AGAAGAAA?CTTACGAGGA?TT?CCC
CTA?GTA?ACG?GCG?AGT
(7) ITS2 of the heavy lip stem of noble dendrobium and 5.8S and 28S rrna part rDNA sequence:
AA?CCAT?CGA?GTC?TTT?GAA?CGC?A
AGTTGCGCCTG?AGGCCAACCG?GCTGAGGGCA
CGTCCGCC?TGGGCGTC?AAGCATTT?TATCGCTC?CGTGCCTA
GTCTCCCA?TCCATGGA?TGTGTTGC?CAAGGCTC?GGATGTGC
ACGGTGGC?TCGTCGTG?CCCCTTGG?TGCGGCGG?GCTGAAGG
GCGGGTCA?TCTTCTCG?TTGGCTGC?CAACAATA?AGGGGTGG
ATTAAATA?AGGCCTAT?GCTATTGT?GTCAAGCG?CGCCTGAG
AGATGGTC?ATACTTAT?TAGGTGAT?CCCAATTC?ATGCGTCG
ATCCATGG?ATGGCGTA?TCGAATGT?GACCCCAG?GATGGGCG
AGGCCACC?CGCCGAGT?TTAAGCAT?ATCAATAA?GCGGAGGA
GAAGAAAC?TTA?CGA?GGA?TT?CCC?CTA?GTA?ACG?GCG?AGT
(8) ITS2 of little Dendrobium tosaense Makino and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTGCGCCC?GAGGCC
AATCGGCC?AAGGGCAC?GTTCGCCTG?GGCGTCAA?GCATTACG
TCGCTCCA?TGTCAACTC?CATCC?CTTC?TTTTGATG?GGCTGGTA
AAAGGCTT?GGACGTGT?AAAGTGGC?TCGTCGTG?CCCCTCGG
TGCGGCGG?GCTTAAGA?GCGGGTCAT?CATCTCGT?TGGCTACG
AACAATTA?CGTGTGGA?TTAAAGTG?CGGCCTAT?CTTATTGT
GTCATGTA?TGCCTGAG?AAGATTAT?GCATTTTT?AGGTGATC
CCAAATCA?TGCGTCGA?TCCATGGA?TGACGTTT?GGAATGCG
ACCCCAGG?ATGGGCGA?GGTCACCC?GCTGAGTT?TAAGCATA
TCAATAAG?CGGAGGAG?AAGAAACT?TACGAGGA?TTCCCCTA
GTAACG?GCG?AGT
(9) ITS2 of the poly-stem of noble dendrobium and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTTG?CGCCCGAGGC
CAATCGGC?CAAGGGCA?CGTTCGCC?TGGGCGTC?AAGCATTA
TGACGCTC?CCTGTCAA?CTCCATCC?CATCTTTT?AATGGGCT
GGCGAAGG?CTCGGATG?TGCAAAGT?GGCTCGTC?GTGCCTCT
CAGTGCGG?CGGGCTTA?AGAGCGGG?TCATCATC?TCGTTGGC
TACGAACA?ATTACGGG?TGGATTAA?AGTGAGGC?CTATATTA
TATTGTCG?TGTATGCC?TGAGAGAA?GATTATTC?ATTTTTTG
GTTATCCC?AAATCATG?CGTCGATC?CATAGATC?ACGTTTGG
AATGCGAC?CCCAGGAT?GGGCGAGG?TCACCCGC?TGAGTTTA
AGCATATC?ACTAAGCG?GAGGAGAA?GAAACTTA?CGAGGATT
CCC?CTA?GTA?ACG?GCG?AGT
(10) ITS2 of the U.S. flower stem of noble dendrobium and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?A?AGTTGCGC?CCGAGGCC
AACCGGCC?AAGGGCAC?GTCCGCCT?GGGCGTCA?GTCATTTT
ATCGCTCT?GTGCCAAG?CTACCCAT?CGATTGAT?GTGTTGGC
GAAGGCTT?GGATGTGC?ATTATGGC?TCATCGTG?CCCCTGGG
CGCGGCGG?GCTGAAGA?GCGGGTCA?TCATCTCG?TTGGCTGC
GAACAATA?AGGGGTGG?ACTGAAGG?GAGACCTA?TGCTATTG
TGTCGTGC?ATGCCTAA?GAGTTGAT?CATACCTT?TGAGGAGA
TCCCAAAT?CATGCGTC?GATCTATG?GACGGCGC?TTTGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CCGCCGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AGAAGAAA?CTTACGAG?GATTCCCC
TAGTAACG?GCGAGT
(11) ITS2 of sieve river stem of noble dendrobium and 5.8S and 28S rrna part rDNA sequence:
AACCATCG?AGTCTTTG?AACGCAAG?TTGCGCCC?GAGGCCAA
CCGGCCAA?GGGCACGT?CTGCCTGG?GCGTCAAG?CATTACAT
CGCTCCAT?GTCAAGTC?ACCCTTCG?ATGGATGG?GCTGGCGA
AGGCTTGG?ATGTGCAC?AGTGGCTC?GTCGTGCC?ACTTGGTG
CGGCGGGC?TGAAGAGC?GGGTCGTC?ATCCTCTT?GGCCGCGA
ACAATAAT?GGGTGGAT?TAAAGCGA?TGCCTAAG?TTATTGTG
TGTGCATG?CCTAAGAG?ATGATTAT?ACTTTTTA?AGTGATCC
CAATTCAT?GCGTCGAT?CCATGGAT?GACGTTTT?GAATGCGA
CCCCAGGA?TAGGCGAG?GCCACCCG?CCGAGTTT?AAGCATAT
CAATAAGC?GGAGGAGA?AGAAACTT?ACGAGGAT?TCCCCTAG
TAACGGCG?AGT
(12) ITS2 of dendrobium moniliformeSweet and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTT?GCGCCTGA?GGCC
AACCGGCT?GAGGGCAC?GTCCGCCT?GGGCGTCA?AGCATTTT
ATCACTCC?GTGCCTAC?TCTCCCAT?CCATGGAT?GTGTTGCT
AAGGCTCG?GATGTGCA?CGGTGGCT?CGTCGTGC?CCCTTGGT
GCGGCGGG?CTGAAGGG?CGGGTCAT?CTTCTCGT?TGGCTGCC
AACAATAA?GGGGTGGA?TTAAATAA?GGCCTATG?CTATTGTG
TCAAGCGC?GCCCGAGA?GATGGTCA?TACTTTTT?AGGTGATC
CCAATTCA?TGCGTTGA?TCCATGGA?TGGCGTAT?CGAATGTG
ACCCCAGG?ATGGGCGA?GGCCACCC?GCTGAGTT?TAAGCATA
TCAATAAG?CGGAGGAG?AAGAAACT?TACGAGGA?TT?CCC?CTA
GTA?ACG?GCG?AGT
(13) ITS2 of the spoon lip stem of noble dendrobium and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTTGAACGCA?AGTTGCGC?CTGAGGCC
AACCGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGCGATAT
GTCGCTCT?GTGCTAAG?TTAACCAT?CGATGGAT?GGGTTGGC
GAAGGCTC?GGATGTGC?ATAATGGC?TCGTTGTG?CCCCTCAG
CGTGGCGG?GCTGAAGA?GGGGGTCA?TCATCTTG?TTAGTCGC
AAACAATA?AGGGGTGG?ATAAATAT?GAGGCCTA?TGTTATTG
TGTTGTGC?ATGCCCAA?GAGATGAT?TAAACCTT?TTTTGTGA
TCCTAAAT?CATGCGTC?AATCCATG?TATGGCGT?TTTGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CCGCCGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AGAAGAAA?CTTACAAG?GATTCCCC
TAGTAACG?GCGAGT
(14) ITS2 of dendrobium stem and 5.8S and 28S rrna part rDNA sequence:
AACCAT?CGA?GTC?TTT?GA?ACGCAAGTTG?CGCCTGAG?GCCA
ATCGGTTG?AGGGCACG?TCCGCCTG?GGCGTCAA?GCATTTTA
TCGCTCCG?TGCCTAGT?CTCCCATC?CATGGATG?TGTTGCCA
AGGCTCGG?ATGTGCAC?GGTGGCTC?GTCGTGCC?CGTTGGTG
CGGCGGGC?TGAAGGGC?GGGTCATC?TTCTCGTT?GGTTGCCA
ACAATAAG?GGGTGGAT?TAAATAAG?GCCTATGC?TATTGTGT
CAAGCGCG?CCTGAGAG?ATGGTCAT?ACTTTTTA?GGTGATCC
CAATTCAT?GCGTCGAT?CCATGGAT?GGCGTATC?GAATGTGA
CCCCAGGA?TGGGCGAG?GCCACCCG?CCGAGTTT?AAGCATAT
CAATAAGC?GGAGGAGA?AGAAACT?TACGAGGA?TTCCCCTA
GTAACGGC?GAGT
(15) ITS2 of Herba Dendrobii and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCCAAGGC
CAACCGGC?TAAGGGCA?CGTCCGCC?TGGGCGTC?AAGCATTT
TATCTCTC?CGTGCCTA?ATCTCCCA?TCCATGGA?TGTGTTAC
TAAGGCTC?GGATGTGC?ATGGTGGC?TCCTCGTG?CCCCTTGG
TGCGGCGG?GCTGAAGG?GCGGGTCA?TCTTCTCG?TTGGTTGC
CAACAATA?AGGGGTGG?ATTAAAAA?AGGCCTAT?GCTATTGT
GATAAGCG?CGCCCGAG?AGATGATC?ATACTTTT?TAGGTGAT
CCCAATCC?ATGCGCTA?ATCCATGG?ATGGCGTA?TCGAATGC
GACCCCAG?GATGGGCG?AGGCCACC?TGCTGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?AAAGAAAC?TTACGAGG?ATTCCCCT
AGTAACGGCGAGT
(16) ITS2 of Dendrobium williamsonii Day et Reichb. F. and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AA?GTT?GCG?CCT?GAG?GCC
AACCGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGCGTTAC
GTCACTCC?GTGCCAAC?TCCATTCC?ATCGATGG?ATGGGCCG
GTGAAGGC?TCGGATGT?GTAGAGTG?GCTCGTCG?TGCCCGTC
GGTGCGGC?GGGTTGAA?GAGTGGGT?CATCATCT?CGTTGGCT
GCGAACAA?TAAGGGGT?GGAATAAA?GTGAGACC?TATGTTAT
TGTGTC?GT?GTATGCCT?GAGAGAAG?ACCATTAT?CAGGAGAT
CCCAAATC?ATGCGTCG?ATCCATGA?ATGGCGTT?TGGAATGC
GACCCCAG?GATGGGCG?AGGCCACC?CGCCGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?GAAGAAAC?TTACGAGG?ATTCCCCT
AGTAACGG CGAGT; Or
(17) any has the dna sequence dna of at least 98.78% homology to sequence (16) with above-mentioned sequence (1).
2. the described dna sequence dna of claim 1, wherein said and above-mentioned sequence (1) any dna sequence dna with at least 98.78% homology to the sequence (16) is selected from:
(1a) ITS2 of hooked dendrobium and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTGCGCCC
GAGGCCAA?CCGGCTAA?GGGCACGT?CCGCCTGG?GCGTCAAG
CATTTTAT?CGCTCCGT?GCCTAATCTCCCATCC?ATGTATGTGT
TGCAAAGG?CTCGGATG?TGCACGGT?GGCTCCTC?GTGCCCCT
TGGTGCGG?CGGGCTGA?AGGCCGGG?TCATCTTC?TCGTTGGC
TGCCAACA?ATAAGGGG?TGGATTAA?ACAAGGCC?TATGCTG
TTGTGATA?TGCGCGCC?CGAGAGAT?GATCATAC?TTTTTAGG
TGATCCCA?ATTCATGC?GCTGATCC?ATGGATGG?CGTATCGA
ATGCGACC?CCAGGATG?GGCGAGGC?CACCCGCT?GAGTTTAA
GCATATCAAT?AAG?CGGAGG?AAAA?GAAA?CTTACGAGGATT
CCC?CTA?GTA?ACG?GCG?AGT
(1b) ITS2 of hooked dendrobium and 5.8S and 28S rrna part rDNA sequence:
AACCAT?CGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCCGAGGC
CAACCGGC?TAAGGGCA?CGTCCGCC?TGGGCGTC?AAGCATTT
TATCGCTC?CGTGCCTA?ATCTCCCA?TCCATGTA?TGTGTTGC
AAAGGCTC?GGATGTGC?ACGGTGGC?TCCTCGTG?CCCCTTGG
TGCGGCGG?GCTGAAGG?CCGGGTCA?TCTTCTCG?TTGGTTGC
CAACAATA?AGGGGTGG?ATTAAACA?AGGCCTAT?GCTGTTGT
GATATGCG?CGCCCGAG?AGATGATC?ATACTTTT?TAGGTGAT
CCCAATTC?ATGCGCTG?ATCCATGG?ATGGCGTA?TCGAATGC
GACCCCAG?GATGGGCG?AGGCCACC?CGCTGAGT?TTAAGCAT
ATCAATAA?GCGG?AGG?AAAA?GAAA?CTTA?CGA?GGATT?CCC
CTA?GTA?ACG?GCG?AGT
(2a) ITS2 of Dendrobium chrysanthum and 5.8S and 28S rrna part rDNA sequence:
AACCATCGA?GTC?TTT?GAA?CGC?AA?GTTGCGCC?CGAGGCCA
ACCTGCCA?AGGGCACG?TCTGCCTG?GGCGTCAA?GCATTTTG
TTGCTCCG?TGTCTCGT?CACCCATC?AATGGATG?TGCTGGCG
AAGGCTCG?GATGTGCA?CGGTGGCC?CGTCGTGC?CCCTCGGT
GCGGCGGG?CTTAAGAG?CGGGTCGC?CATCTCGT?TGGTTGCG
AACAATAA?GGGGTGGA?TTAAATGG?AGGCCTAT?GCTATTGT
GTCGTGCA?TGCCTAAG?AGATTAAA?TTTTTTAG?GTGATCCC
AAATCACG?CGTCGATC?CACGGATG?ATGCTTTG?AATGCGAC
CCCAGGAT?GGGCGAGG?CCACCCGC?CGAGTTTA?AGCATATC
AATAAGCG?GAGGAGAA?GAAACTTA?CAAGGATT?CCCCTAGT
AACGGCG?AGT
(3a) ITS2 of Dendrobium chrysotoxum and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCCGAGGC
CAATCGGC?CAAGGGCA?CGTCTGCC?TGGGCGTC?AAGCGTTA
CGTCGCTC?CGTGTCCA?GCCACCCG?TCGATGGA?TGGGCTGG
CGATGGCT?CGGATGTG?CACAGTGG?CTCCTCGT?GCCCCTCG
GCACGGCG?GGCTTAAG?AGCGGGTA?ATCATCTC?GTTGGCTG
CGAACAAT?ATGGGGTG?GATAAATG?CGATGCCT?ATGATATT
GTGCCGTG?CATGCCCG?AGAGATGA?TCATACTT?ATTCAGGT
GATCCCAA?ACCATGCG?TCGATCCA?TGGATGGC?GTTTTGAA
TGCGACCC?CAGGACAG?GCGAGGCC?ACCCGCCG?AGTTTAAG
CATATCAA?TAAGCGGA?GGAGAAGA?AACTTACG?AGGATT
CCC?CTA?GTA?ACG?GCG?AGT
(3b) ITS2 of Dendrobium chrysotoxum and 5.8S and 28S rrna part rDNA sequence:
AACCAT?CGA?GTC?TTT?GAA?CGC?AAG?TTGCGCCC?GAGGCCAA
TCGGCCAA?GGGCACGT?CTGCCTGG?GCGTCAAG?CGTTACGT
CGCTCCGT?GTCCAGCC?ACCCGTCG?ATGGATGG?GCTGGCGA
TGGCTCGG?ATGTGCAC?AGTGGCTC?CTCGTGCC?CCTCGGCA
CGGCGGGC?TTAAGAGC?GGGTAATC?ATCTCGTT?GGCTGCGA
ACAATATG?GGGTGGAT?AAATGCGA?TGCTTATG?ATATTGTG
CCGTGCAT?GCCCGAGA?GATGATCA?TACTTATT?CAGGTGAT
CCCAAACC?ATGCGTCG?ATCCATGG?ATGGCGTT?TTGAATGC
GACCCCAG?GACAGGCG?AGGCCACC?CGCCGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?GA?AGA?AAC?TTA?CGA?GGATT?CCC
CTA?GTA?ACG?GCG?AGT
(4a) ITS2 of Dendrobium crepidatum lindl et paxt and 5.8S and 28S rrna part rDNA sequence:
AACCAT?CGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCCGAGGC
CAATCGGC?CAAGGGCA?CGTCTGCC?TGGGCGTC?AAGCATTT
TGTTGCTC?CGTGTCTA?CTCACACC?CATCGATG?GATGTGCT
GGTGTAGG?CTCGGATG?TGCATGCT?GGCTCGTC?GTTCTCCC
AGGTGCGG?CGGGCTGA?AGAGCGGG?TCATCCTC?TCGTTTGC
GGCGAACA?ATAAGGGG?TGGATTGA?AGAGAGGT?CTATGCTA
TTGTGTCG?TGCATGCC?TGAGAGAT?GATTAACT?TTTTTGTG
ATCCCAAA?TCATGCAT?CGATCCAT?GGATGATG?CTTTGAAT
GTGACCCC?AGGATGGG?CGAGGCTA?CCCGCTGA?GTTTAAGC
ATATCAAT?AAGCGGAG?GAGAAGAA?ACTTACGA?GGATTCCC
CTAGTAAC?GGCGAGT
(4b) ITS2 of Dendrobium crepidatum lindl et paxt and 5.8S and 28S rrna part rDNA sequence:
AACCAT?CGA?GTC?TTT?GAA?CGC?AAGTTG?CGCCCGAG
GCCAATCG?GCCAAGGG?CACGTCTG?CCTGGGCG?TCAAGCAT
TTTGTTGC?TCCGTGTC?TACTCACA?CCCATCGA?TGGATGTG
CTGGTGTA?GGCTCGGA?TGTGCATG?CTGGCTCG?TCGTTCTC
CCAGGTGC?GGCGGGCT?GAAGAGCG?GGTCATCC?TCTCGTTT
GCGGCGAA?CAATAAGG?GGTGGATT?GAAGAGAG?GTCTATGC
TATTGTGT?CGTGCATG?CCTGAGAG?ATGATTAA?CTTTTTTG
TGATCCCA?AATCATGC?ATC?GATC?CATGGATG?ATGC?TTTG
AATG?TGAC?CCCA?GGAT?GGGC?GAGG?CTAC?CCGC?TGAG
TTTA?AGCA?TATC?AATA?AGCG?GAGG?AGAA?GAAA?CTTA
CGAG?GA?TT?CCC?CTA?GTA?ACG?GCG?AGT
(5a) ITS2 of dendrobium and 5.8S and 28S rrna part rDNA sequence:
AACCAT?CGA?GTC?TTT?GAA?CGC?AAG?TTGCGCCC?GAGGCCAA
CCGGCCAA?GGGCACGT?CTGCCTGG?GCGTCAAG?CGTTATGT
CGCTCCGT?GTTAACTC?CATCCCAT?CGATGTAT?GGGCTGGC
GAAGGCTC?GGATGTGC?AGTGTGGC?TCGTCGTG?CCCCTTGG
TGCGGCGA?GCTGAAA?GCGGGTAA?TCATCTCG?TTGGCTGC
GAACGATA?AGGGGTGG?ATTAAAGC?GAGACCTA?TGTTATTG
TGTCGTGT?ATGCCCGA?GAGAAGAT?TATACATA?CTCAGGTG
ATCCTGAA?TCATGCGT?CGATCAAA?GGACGGCG?CTTGGAAT
GCGACCCC?AGGATGGG?CGAGGCCA?CCCGCCGA?GTTTAAGC
ATATTAAT?AAGCGGAG?GAGAAGAA?ACTTACGA?GGATTCCC
CTA?GTA?ACG?GCG?AGT
(5b) ITS2 of dendrobium and 5.8S and 28S rrna part rDNA sequence: AAC CATCGA GTC TTT GAA CGC AAGTTGCGC CCGAGGCCAA
CCGGCCAA?GGGCACGT?CTGCCTGG?GCGTCAAG?CGTTATGT
CGCTCCGT?GTTAACTC?CATCCCAT?CGATGTAT?GGGCTGGC
GAAGGCTC?GGATGTGC?AGTGTGGC?TCGTCGTG?CCCCTTGG
TGCGGCGA?GCTGAAAA?GCGGGTAA?TCATCTCG?TTGGCTGC
GAACGATA?AGGGGTGG?ATTAAAGC?GAGACCTA?TGTTATTG
TGTCGTGT?ATGCCCGA?GAGAAGAT?TATACATA?CTCAGGTG
ATCCTGAA?TCATGCGT?CGATCAAA?GGACGGCG?CTTGGAAT
GCGACCCC?AGGATGGG?CGAGGCCA?CCCGCCGA?GTTTAAGC
ATATTAAT?AAGCGGAG?GAGAAGAA?ACTTACGA?GGATTCCC
CTAGTAAC?GGCGAGT
(6a) ITS2 of Dendrobium fimbriatum Hook. and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTGCGCCCGA
GGCCAACA?GGCCAAGG?GCACGTCT?GCCTGGGC?GTCAAGTG
TTATGTCG?CTCCATGC?CAAGTCAC?CCATCGAT?GGATGGGC
CGGCGAAG?GCTCGGAT?GTGCACAG?TGGCTCGC?CGTGCCCC
TCGGTGCG?GCGGGCTG?AAGAGCGG?GTCATTAT?CTTGTTGG
CTGCAGGC?AATAATGG?GTGGATAA?ACGCGAGG?CCTTTGTT
ATTGTGTC?GTGCATGC?CTAAGATA?TGATTAAA?CTTTTTAG
GTGATCCC?AAATCATG?CGTCGATC?CACGGATG?GCGTTTTG
AATGCGAC?CCCAGGAT?GGGCGAGG?CCACCCGC?CGAGTTTA
AGCATATC?AATAAGCG?GAGGAGAA?GAAACTTA?CGA?GGA
TT?CCC?CTA?GTA?ACG?GCG?AGT
(6b) ITS2 of Dendrobium fimbriatum Hook. and 5.8S and 28S rrna part rDNA sequence: AAC CATCGA GTC TTT GAA CGC AAGTTGCG CC CGAGGCCA
ACAGGCCA?AGGGCACG?TCTGCCTG?GGCGTCAA?GTGTTATG
TCGCTCCA?TGCCAAGT?CACCCATC?GATGGATG?GGCCGGCG
AAGGCTCG?GATGTGCA?CAGTGGCT?CGCCGTGC?CCCTCGGT
GCGGCGGG?CTGAAGAG?CGGGTCAT?TATCTTGT?TGGCTGCA
GGCAATAA?TGGGTGGA?TAAACGCG?AGGCCTTT?GTTATTGT
GTCGTGCA?TGCCTAAG?ATATGATT?AAACTTTT?TAGGTGAT
CCCAAATC?ATGCGTCG?ATCCACGG?ATGGCGTT?TTGAATGC
GACCCCAG?GATGGGCG?AGGCCACC?CGCCGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?GAAGAAAC?TTACGAGGA?TT?CCC
CTA?GTA?ACG?GCG?AGT
(7a) ITS2 of the heavy lip stem of noble dendrobium and 5.8S and 28S rrna part rDNA sequence:
AACCAT?CGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCTGAGGC
CAACCGGC?TGAGGGCA?CGTCCGCC?TGGGCGTC?AAGCATTT
TATCGCTC?CGTGCCTA?GTCTCCCA?TCCATGGA?TGTGTTGC
CAAGGCTC?GGATGTGC?ACGGTGGC?TCGTCGTG?CCCCTTGG
TGCGGCGG?GCTGAAGG?GCGGGTCA?TCTTCTCG?TTGGCTGC
CAACAATA?AGGGGTGG?ATTAAATA?AGGCCTAT?GCTATTGT
GTCAAGCG?CGCCTGAG?AGATGGTC?ATACTTAT?TAGGTGAT
CCCAATTC?ATGCGTCG?ATCCATGG?ATGGCGTA?TCGAATGT
GACCCCAG?GATGGGCG?AGGCCACC?CGCCGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?GAAGAAAC?TTACGAGG?ATTCCCCT
AGTA?ACG?GCG?AGT
(8a) ITS2 of little Dendrobium tosaense Makino and 5.8S and 28S rrna part rDNA sequence:
AACCAT?CGA?GTC?TTT?GAA?CGC?AAG?TTGCGCCC?GAGGCCAA
TCGGCCAA?GGGCACGT?TCGCCTGG?GCGTCAAG?CATTACGT
CGCTCCAT?GTCAACTC?CATCCCTT?CTTTTGAT?GGGCTGGT
AAAAGGCT?TGGACGTG?TAAAGTGG?CTCGTCGT?GCCCCTCG
GTGCGGCG?GGCTTAAG?AGCGGGTC?ATCATCTC?GTTGGCTA
CGAACAAT?TACGTGTG?GATTAAAG?TGCGGCCT?ATCTTATT
GTGTCATG?TATGCCTG?AGAAGATT?ATGCATTT?TTAGGTGA
TCCCAAAT?CATGCGTC?GATCCATG?GATGACGT?TTGGAATG
CGACCCCA?GGATGGGC?GAGGTCAC?CCGCTGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AGAAGAAA?CTTACGAG?GATTCCCC
TAGTAACG?GCGAGT
(9a) ITS2 of the poly-stem of noble dendrobium and 5.8S and 28S rrna part rDNA sequence:
AACCATCGAGTC?TTT?GAA?CGC?AAG?TTGCGCCC?GAGGCCAA
TCGGCCAA?GGGCACGT?TCGCCTGG?GCGTCAAG?CATTATGA
CGCTCCCT?GTCAACTC?CATCCCAT?CTTTTAAT?GGGCTGGC
GAAGGCTC?GGATGTGC?AAAGTGGC?TCGTCGTG?CCTCTCAG
TGCGGCGG?GCTTAAGA?GCGGGTCA?TCATCTCG?TTGGCTAC
GAACAATT?ACGGGTGG?ATTAAAGT?GAGGCCTA?TATTATAT
TGTCGTGT?ATGCCTGA?GAGAAGAT?TATTCATT?TTTTGGTT
ATCCCAAA?TCATGCGT?CGATCCAT?AGATCACG?TTTGGAAT
GCGACCCC?AGGATGGG?CGAGGTCA?CCCGCTGA?GTTTAAGC
ATATCACT?AAGCGGAG?GAGAAGAA?ACTTACGA?GGATTCCC
CTA?GTA?ACG?GCG?AGT
(10a) ITS2 of the U.S. flower stem of noble dendrobium and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTT?GCGC?CCGAGGC
CAACCGG?CCAAGGGC?ACGTCCGC?CTGGGCGT?CAGTCATT
TTATCGCT?CTGTGCCA?AGCTAACC?ATCGATTG?ATGTGTTG
GCGAAGGC?TTGGATGT?GCATTATG?GCTCATCG?TGCCCCTG
GGCGCGGC?GGGCTGAA?GAGCGGGT?CATCATCT?CGTTGGCT
GCGAACAA?TAAGGGGT?GGACTGAA?GGGAGAC?CTATGCTA
TTGTGTCG?TGCATGCC?TAAGAGTT?GATCATAC?CTTTGAGG
AGATCCCA?AATCATGC?GTCGATCT?ATGGACGG?CGCTTTGA
ATGCGACC?CCAGGATG?GGCGAGGC?CACCCGCC?GAGTTTAA
GCATATCA?ATAAGCGG?AGGAGAAG?AAACTTAC?GAGGATTC
CC?CTA?GTA?ACG?GCG?AGT
(10b) ITS2 of the U.S. flower stem of noble dendrobium and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTGCGCCC?GAGGCC
AACCGGCC?AAGGGCAC?GTCCGCCT?GGGCGTCA?GTCATTTT
ATCGCTCT?GTGCCAAG?CTACCCAT?CGATTGAT?GTGTTGGC
GAAGGCTT?GGATGTGC?ATTATGGC?TCATCGTG?CCCCTGGG
CGCGGCGG?GCTGAAGA?GCGGGTCA?TCATCTCG?TTGGCTGC
GAACAATA?AGGGGTGG?ACTGAAGG?GAGACCTA?TGCTATTG
TGTCGTGC?ATGCCTAA?GAGTTGAT?CATACCTT?TGAGGAGA
TCCCAAAT?CATGCGTC?GATCTATG?GACGGCGC?TTTGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CCGCCGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AGAAGAAA?CTTACGAG?GATTCCCC
TAGTAACGGCG?AGT
(11a) ITS2 of dendrobium moniliformeSweet and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGT?TGCGCCTG?AGGCCA
ACCGGCTG?AGGGCACG?TCCGCCTG?GGCGTCAA?GCATTTTA
TCACTCCG?TGCCTACT?CTCCCATC?CATGGATG?TGTTGCTA
AGGCTCGG?ATGTGCAC?GGTGGCTC?GTCGTGCC?CCTTGGTG
CGGCGGGC?TGAAGGGC?GGGTCATC?TTCTCGTT?GGCTGCCA
ACAATAAG?GGGTGGAT?TAAATAAG?GCCTATGC?TATTGTGT
CAAGCGCG?CCCGAGAG?ATGGTCAT?ACTTTTTA?GGTGATCC
CAATTCAT?GCGTTGAT?CCATGGAT?GGCGTATC?GAATGTGA
CCCCAGGA?TGGGCGAG?GCCACCCG?CTGAGTTT?AAGCATAT
CAATAAGC?GGAGGAGA?AGAAACTT?ACGAGGAT?TCCCCTAG
TAACGGCG?AGT
(11b) ITS2 of dendrobium moniliformeSweet and 5.8S and 28S rrna part rDNA sequence:
AACCATCG?A?GTC?TTT?GAA?CGC?AAGTTGCGC?CTGAGGCCAA
CCGGCTGA?GGGCACGT?CCGCCTGG?GCGTCAAG?CATTTTAT
CACTCCGT?GCCTACTC?TCCCATCC?ATGGATGT?GTTGCTAA
GGCTCGGA?TGTGCACG?GTGGCTCG?TCGTGCCC?CTTGGTGC
GGCGGGCT?GAAGGGCG?GGTCATCT?TCTCGTTG?GCTGCCAA
CAATAAGG?GGTGGATT?AAATAAGG?CCTATGCT?ATTGTGTC
AAGCGCGC?CCGAGAGA?TGGTCATA?CTTTTTAG?GTGATCCC
AATTCATG?CGTTGATC?CATGGATG?GCGTATCG?AATGTGAC
CCCAGGAT?GGGCGAGG?CCACCCGC?TGAGTTTA?AGCATATC
AATAAGCG?GAGGAGAA?GAAACTTA?CGAGGATT?CCCCTAGT
AACGGCGAGT
(12a) ITS2 of the spoon lip stem of noble dendrobium and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGT?TGCGCCTGAG?GCC
AACCGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGCGA?TAT
GTCGCTCT?GTGTTAAG?TTAACCAT?CGATGGAT?GGGTTGGC
GAAGGCTC?GGATGTGC?ATAATGGC?TCGTTGTG?CCCCTCAG
CGTGGCGG?GCTGAAGA?GGGGGTCA?TCATCTTG?TTAGTCGC
AAACAATA?AGGGGTGG?ATAAATAT?GAGGCCTA?TGTTATTG
TGTTGTGC?ATGCCCAA?GAGATGAT?TAAACCTT?TTTTGTGA
TCCTAAAT?CATGCGTC?AATCCATG?TATGGCGT?TTTGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CCGCCGAG?TTTAAGC
ATATCAAT?AAGCGGTG?GAGAAGAA?ACTTACAA?GGATTCCC
CTAGTAAC?GGCGAGT
(12b) ITS2 of the spoon lip stem of noble dendrobium and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCTGAGGC
CAACCGGC?CAAGGGCA?CGTCTGCC?TGGGCGTC?AA?GCGATA
TGTCGCTC?TGTGCTAA?GTTAACCA?TCGATGGA?TGGGTTGG
CGAAGGCT?CGGATGTG?CATAATGG?CTCGTTGT?GCCCCTCA
GCGTGGCG?GGCTGAAG?AGGGGGTC?ATCATCTT?GTTAGTCG
CAAACAAT?AAGGGGTG?GATAAATA?TGAGGCCT?ATGTTATT
GTGTTGTG?CATGCCCA?AGAGATGA?TTAAACCT?TTTTTGTG
ATCCTAAA?TCATGCGT?CAATCCAT?GTATGGCG?TTTTGAAT
GCGACCCC?AGGATGGG?CGAGGCCA?CCCGCCGA?GTTTAAGC
ATATCAAT?AAGCGGAG?GAGAAGAA?ACTTACAA?GGATTCCC
CTAGTAAC?GGCGAGT
(13a) ITS2 of dendrobium stem and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTGCGCCT?GAGGCC
AACCGGCT?GAGGGCAC?GTCCGCCT?GGGCGTCA?AGCATTTT
ATCGCTCC?GTGCCTAG?TCTCCCAT?CCATGGAT?GTGTTGCC
AAGGCTCG?GATGTGCA?CGGTGGCT?CGTCGTGC?CCCTTGGT
GCGGCGGG?CTGAAGGG?CGGGTCAT?CTTCTCGT?TGGCTGCC
AACAATAA?GGGGTGGA?TTAAATAA?GGCCTATG?CTATTGTG
TCAAGCGC?GCCTGAGA?GATGGTCA?TACTTATT?AGGTGATC
CCAATTCA?TGCGTCGA?TCCATGGA?TGGCGTAT?CGAATGTG
ACCCCAGG?ATGGGCGA?GGCCACCC?GCCGAGTT?TAAGCATA
TCAATAAG?CGGAGGAG?AAGAAACT?TACGAGGA?TTCCCCTA
GTAACGGC?GAGT
(13b) ITS2 of dendrobium stem and 5.8S and 28S rrna part rDNA sequence:
AACCATCGA?GTC?TTT?GAA?CGC?AAG?TTGCGCCT?GAGGCCAA
CCGGCTGA?GGGCACGT?CCGCCTGG?GCGTCAAG?CATTTTAT
CGCTCCGT?GCCTAGTC?TCCCATCC?ATGGATGT?GTTGCCAA
GGCTCGGA?TGTGCACG?GTGGCTCG?TCGTGCCC?CTTGGTGC
GGCGGGCT?GAAGGGCG?GGTCATCT?TCTCGTTG?GCTGCCAA
CAATAAGG?GGTGGATT?AAATAAGG?CCTATGCT?ATTGTGTC
AAGCGCGC?CTGAGAGA?TGGTCATA?CTTATTAG?GTGATCCC
AATTCATG?CGTCGATC?CATGGATG?GCGTATCG?AATGTGAC
CCCAGGAT?GGGCGAGG?CCACCCGC?CGAGTTTA?AGCATATC
AATAAGCG?GAGGAGAA?GAAACTTA?CGAGGATT?CCCCTAGT
AACGGCGAGT
(14a) ITS2 of Herba Dendrobii and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCCAAGGC
CAACCGGC?TAAGGGCA?CGTCCGCC?TGGGCGTC?AAGCATTT
TATCTCTC?CGTGCCTA?ATCTCCCA?TCCATGGA?TGTGTTAC
TAAGGCTC?GGATGTGC?ATGGTGGC?TCCTCGTG?CCCCTTGG
TGCGGCGG?GCTGAAGG?GCGGGTCA?TCTTCTCG?TTGGTTG
CCAACAAT?AAGGGGTG?GATTAAAA?AAGGCCTA?TGCTATTG
TGATAAGC?GCGCCCGA?GAGATGAT?CATACTTT?TTAGGTGA
TCCCAATC?CATGCGCT?AATCCATG?GATGGCGT?ATCGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CTGCTGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AAAAGAAA?CTTACGAG?GATTCCCC
TAGTAACG?GCGAGT
(14b) ITS2 of Herba Dendrobii and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCCAAGGC
CAACCGGC?TAAGGGCA?CGTCCGCC?TGGGCGTC?AAGCATTT
TATCTCTC?CGTGCCTA?ATCTCCCA?TCCATGGA?TGTGTTAC
TAAGGCTC?GGATGTGC?ATGGTGGC?TCCTCGTG?CCCCTTGG
TGCGGCGG?GCTGAAGG?GCGGGTCA?TCTTCTCG?TTGGTTGC
CAACAATA?AGGGGTGG?ATTAAAAA?AGGCCTAT?GCTATTGT
GATAAGCG?CGCCCGAG?AGATGATC?ATACTTTT?TAGGTGAT
CCCAATCC?ATGCGCTA?ATCCATGG?ATGGCGTA?TCGAATGC
GACCCCAG?GATGGGCG?AGGCCACC?TGCTGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?AAAGAAAC?TTACGAGG?ATTCCCCT
AGTAACGG?CGAGT
(14c) ITS2 of Herba Dendrobii and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAGTTGCG?CCCAAGGC
CAACCGGC?TAAGGGCA?CGTCCGCC?TGGGCGTC?AAGCATTT
TATCTCTC?CGTGCCTA?ATCTCCCA?TCCATGGA?TGTGTTAC
TAAGGCTC?GGATGTGC?ATGGTGGC?TCCTCGTG?CCCCTTGG
TGCGGCGG?GCTGAAGG?GCGGGTCA?TCTTCTCG?TTGGTTGC
CAACAATA?AGGGGTGG?ATTAAAA AGGCCTAT?GCTATTGT
GATAAGCG?CGCCCGAG?AGATGATC?ATACTTTT?TAGGTGAT
CCCAATCC?ATGCGCTA?ATCCATGG?ATGGCGTA?TCGAATGC
GACCCCAG?GATGGGCG?AGGCCACC?TGCTGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?AAAGAAAC?TTACGAGG?ATTCCCCT
AGTAACGG?CGAGT
(14d) ITS2 of Herba Dendrobii and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCA?AGG
CCAACCGG?CTAAGGGC?ACGTCCGC?CTGGGCGT?CAAGCATT
TTATCTCT?CCGTGCCT?AATCTCCC?ATCCATGG?ATGTGTTA
CTAAGGCT?CGGATGTG?CATGGTGG?CTCCTCGT?GCCCCTTG
GTGCGGCG?GGCTGAAG?GGCGGGTC?ATCTTCTC?GTTGGTTG
CCAACAAT?AAGGGGTG?GATTAAAA?AAGGCCTA?TGCTATTG
TGATAAGC?GCGCCCGA?GAGATGAT?CATACTTT?TTAGGTGA
TCCCAATC?CATGCGCT?AATCCATG?GATGGCGT?ATCGAATG
CGACCCCA?GGATGGGC?GAGGCCAC?CTGCTGAG?TTTAAGCA
TATCAATA?AGCGGAGG?AAAAGAAA?CTTACGAG?GATTCCCC
TAGTAACG?GCGAGT
(14e) ITS2 of Herba Dendrobii and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGC?CCA?AGG
CCAACCGG?CTAAGGGC?ACGTCCGC?CTGGGCGT?CAAGCATT
TTATCTCT?CCGTGCCT?AATCTCCC?ATCCATGG?ATGTGTTA
CTAAGGCT?CGGATGTG?CATGGTGG?CTCCTCGT?GCCCCTTG
GTGCGGCG?GGCTGAAG?GGCGGGTC?ATCTTCTC?GTTGGTTG
CCAACAAT?AAGGGGTG?GATTAAAA?AAGGCCTA?TGCTATTG
TGATAAGC?GCGCCCGAG?AGATGATC?ATACTTTT?TAGGTGAT
CCCAATCC?ATGCGCTA?ATCCATGG?ATGGCGTA?TCGAATGC
GACCCCAG?GATGGGCG?AGGCCACC?TGCTGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?AAAGAAAC?TTACGAGG?ATTCCCCT
AGTAACGG?CGAGT
(15a) ITS2 of Dendrobium williamsonii Day et Reichb. F. and 5.8S and 28S rrna part rDNA sequence:
AAC?CAT?CGA?GTC?TTT?GAA?CGC?AAG?TTG?CGCCTGAG?GCC
AACCGGCC?AAGGGCAC?GTCTGCCT?GGGCGTCA?AGCGTTAC
GTCACTCC?GTGCCAAC?TCCATTCC?ATCGATGG?ATGGGCCG
GTGAAGGC?TCGGATGT?GTAGAGTG?GCTCGTCG?TGCCCGTC
GGTGCGGC?GGGTTGAA?GAGTGGGT?CATCATCT?CGTTGGCT
GCGAACAA?TAAGGGGT?GGAATAAA?GTGAGACC?TATGTTAT
TGTGTCGT?GTATGCCT?GAGAGAAG?ACCATTAT?CAGGAGAT
CCCAAATC?ATGCGTCG?ATCCATGA?ATGGCGTT?TGGAATGC
GACCCCAG?GATGGGCG?AGGCCACC?CGCCGAGT?TTAAGCAT
ATCAATAA?GCGGAGGA?GAAGAAAC?TTACGAGG?ATTCCCCT
AGTAACGG?CGAGT。
3. method of identifying stem of noble dendrobium kind or distinguishing its true and false, this method comprises the steps:
A. from plant sample, extract DNA;
B. the DNA that extracts with step a is a template, with oligonucleotide as polymerase chain reaction (PCR) primer, amplification ITS2 and the section of 5.8S and 28S partly, used primer picks up from the conservative section of plant 5.8S and 28S respectively;
C. use the sequence of the PCR product that direct sequencing technologies determination step b obtained;
D. the comparison of the ITS2 of dna sequence dna that step c is recorded and the arbitrary stem of noble dendrobium kind described in claim 1 or 2 and 5.8S and 28S rrna part rDNA sequence is to determine stem of noble dendrobium kind or to distinguish its true and false.
CNB011024348A 2001-01-31 2001-01-31 DNA sequence of dendrobium and method for discriminating its veriety and judging if it is true or false Expired - Fee Related CN1173989C (en)

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US8442245B2 (en) 2008-12-22 2013-05-14 Siemens Medical Instruments Pte. Ltd. Hearing device with automatic algorithm switching

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CN102732513B (en) * 2012-06-19 2014-01-15 中国医学科学院药用植物研究所 PCR detection method of Dendrobium chrysotoxum by adoption of specific primers
CN103173532B (en) * 2012-09-29 2015-02-18 中国医学科学院药用植物研究所 Method for identifying honeysuckle and lonicera confuse and application of same
CN103571949A (en) * 2013-10-12 2014-02-12 杭州师范大学 Microsatellite molecular markers for identifying hard/soft-foot dendrobium officinale
CN104531872A (en) * 2014-12-29 2015-04-22 杭州师范大学 Molecular specific marker primer and method for identifying dendrobium hercoglossum and dendrobium aduncum
CN106399475B (en) * 2016-08-26 2019-06-04 中国医学科学院药用植物研究所 A method of it obtaining rDNA ITS2 sequence and is used to identify Tiepi Fengdou

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8442245B2 (en) 2008-12-22 2013-05-14 Siemens Medical Instruments Pte. Ltd. Hearing device with automatic algorithm switching

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